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Introduces the research papers to be published in the "European Journal of Biochemistry," succeeding the May 15, 1985. "Primer-independent abortive initiation by wheat-germ RNA polymerase B," by H. Mosig et al; "Protein structure and gene organization of mouse lactate dehydrogenase-A isozyme," S.S.-L. Li et al; "A model for intracellular complexation between gene-5 protein and bacteriophage fd DNA," by G.D. Bayer et al.

The article lists several research papers that would be published in the coming issues of the European Journal of Biochemistry. Some of the articles and topics are related to glucogen synthase, protein phosphates, DNA sequences, etc.

Presents research articles on biochemistry to be published in the "European Journal of Biochemistry." "Secondary and Tertiary Structure Characteristic of Megasphaera elsdenii Flavodoxin in the Reduced State as Determined by Two-Dimensional 1H NMR," by C. P. M. van Mierlo, F. M¨ller, and J. Vervoort; "Presence and Characterization of Glycolipid Sulfotransferase in Human Cancer Serum," by S. Gasa, M. -T. Casl, A. Makita, N. Sakakibara, T. Koyanagi, and T. Atsuta; "Interaction of Hemin With Placental Glutathione Transferase," by A. M. Caccuri; A. Aceto, F. Piemonte, C. di Ilio, N. Rosato, and G. Federici.

Lists articles about biochemistry. "Modulation of the interaction between the two halves of troponin C by the other troponin subunits," by C.-L.A. Wang and J. Gergely; "Sequence determinants of cytosolic N-terminal protein processing," by C. Flinta, B. Persson, H. Jornvall and G. von Heijne; "Effect of benzoate on the metabolism of fructose 2,6-biphosphate in yeast," by J. Francois, E. van Schaftingen and H.-G. Hers; "Isolation and properties of porcine cholesterol acyltransferase," G. Knipping; Others.

Lists the titles of papers that will be published in future issues of the "European Journal of Biochemistry." "Regulation of the ontogeny of rat liver metallothionein mRNA by zinc," by G.K. Andrews et al; "Processing of a plant vacuolar protein precursor in vitro," by T. Hattori et al "Structural studies of the putatitve O-specific polysaccharide of Serratia marcescens 09," by D. Oxley and S.G. Wilkinson.

The article presents a list of forthcoming research papers including "Kinetic Studies of Proton Transfer in the Microenvironment of a Binding Site," by M. Gutman, D. Huppert and E. Nachliel," "The Substrate Binding Site of Aldehyde Reductase From Pig Liver: Stereochemical Investigations Using NADP-2-Oxodiacid Adducts As Probe," by G. Branlant and "Membranes of Protein Bodies: I. Isolation from Cotyledons of Germinating Cucumber Seeds" by U. A. K. Kara and H. Kindl.

Lists articles about biochemistry. "Impact of altered protein structures on the intracellular traffic of a mutated vasopressin precursor from Brattleboro rats," by H. Schmale et al; "Direct electron transfer of redox proteins at the bare classy carbon electrode," by W.R. Hagen; "Molecular cloning of preproacrosin and analysis of its expression pattern in spermatogenesis," by I.M. Adham; "Reduced temperature does not prevent transport of lysosomal integral membrane proteins from endoplasmic reticulum and through the Golgi system to lysosomes," by P.G. Morales; Others.

Offers information on articles for publication in future issues of the "European Journal of Biochemistry". Synthesis of two proteins in chloroplasts and messenger RNA; Types of acids from Mycobacterium thamnopheos; Use of the inhibition of enzymatic antioxidant systems in order to evaluate their physiological importance.

Presents several articles to be published in the "European Journal of Biochemistry." "Modification of Ribosomal Proteins in Sea Urchin Eggs Following Fertilization," by K. Takeshima and E. Nakano; "A Phosphotyrosyl-Protein Phosphatase Activity Associated With Acid Phosphatase From Human Prostate Gland," by H.-C. Li, J. Chernoff, L. B. Chen and A. Kirschonbaum; "Molecular Characterization of the Gene Coding for Major Outer Membrane Protein OmpA from Enterobacter Aerogenes," by G. Braun and S. T. Cole; "Nidogen: A New, Self-Aggregating Basement Membrane Protein," by R. Timpl, M. Dziadek, S. Fujiwara, H. Nowack and G. Wick; Others.

A neurodepressing hormone has been isolated and purified to homogeneity from aqueous extracts of 2000 eyestalks of the Mexican crayfish Procambarus bouvieri (Ortmann). Purification was achieved by gel filtration on Sephadex G-25 and G-15, and preparative paper electrophoresis at four pH values (1.8, 3.6, 6.0 and 10.0). The recovery of hormone activity was 85% and the specific activity was 4.0 × 104 limes that of the starting material. The hormone is a thermostable peptide of approximately 1200 molecular weight and composed of neutral amino acids. No N-terminal group could be found. From its electrophoretic behavior it is concluded that the C-terminal group is also blocked. [ABSTRACT FROM AUTHOR]

Lists papers that will published in the upcoming issues of the "European Journal of Biochemistry". "Eurabutoxin b--Initial protein refinement and sequence analysis at 0.140-nm resolution," by P.E. Bourne and colleagues; "Preferential conformation of substance P in solution," by G. Chassaing and colleagues; "Interaction of actinogelin with actin--No nucleation but high gelation activity".

A series of bacterial cell wall glycopeptides of low molecular weight and cell wall nucleotide precursors have been tested for their inhibitory action on the digestion by T4 lysozyme of a radioactively labeled linear uncrosslinked peptidoglycan. The disaccharide-peptides GlcNAc-MurNAc- L-Ala-D-Glu(A2pm) (C5) and GlcNAc-MurNAc-L-Ala-D-Glu(A2pm-D-Ala) (C6) as well as the monosaccharide-peptide MurNAc-L-Ala-D-Glu(A2pm) were found to be good competitive in- hibitors (with similar Ki values) whereas the disaccharide-pentapeptide GlcNAcMurNAc-L-Ala-D Glu-Gly [This symbol cannot be represented into ASCII Text]-L-Lys-D-Ala was a poor inhibitor. T4 lysozyme did not catalyse transglycosylation reactions from Escherichia coli B peptidoglycan to the disaccharide-peptide C6. No changes were seen in the circular dichroism spectra (200–250 nm) or fluorescence emission spectra upon binding of the good inhibitors. The results obtained indicate that T4 lysozyme has a small active site capable of recognizing a unit consisting of MurNAc-L-Ala-D-Glu(A2pm). [ABSTRACT FROM AUTHOR]

The electron transfer kinetics between the hydrogenase from Desulfovibrio vulgaris (strain Hildenborough) and the mediators methyl viologen, di-(n-aminopropyl) viologen and propyl viologen sulfonate have been investigated by chronoamperometry. Second-order rate constants were calculated on basis of the theory for a simple catalytic mechanism and are compared with the results obtained before by cyclic voltammetry (preceding paper in this journal). From the ionic-strength dependence and the observed differences in the rate constants for the differently charged viologens, the existence of an electrostatic interaction between mediator and a negatively charged part of the protein is confirmed. Chronoamperometry (computer-controlled) was found to possess advantages over cyclic voltammetry in the determination of homogeneous rate constants (faster, more accurate, and better reproducibility). [ABSTRACT FROM AUTHOR]

A purified glycoprotein with blood-group-A specificity, was digested with insoluble pronase to give a serologically active product enriched in peptide-bonded serine and threonine residues; this was degraded with 0.25 M sulphuric in glacial acetic acid for 24 h. The resultant N-acetylgalactosaminyl-peptide were hydrolysed with 0.2 M hydrocholoric acid for 24 h at 110 °C. fractionation of the products on Sephadex G-10, followed by preparative separation on amino-acid analyzer and paper electrophoresis at pH 1.9 yielded the following homogeneous oligopeptides; one tripeptide with the N-terminal sequence Thr-Thr-Ser, three tetrapeptides with the sequences, Thr-Ser-Thr-Ser, Thr-Pro-Thr-Ser and Pro-Thr-Thr-Ser, one pentapeptide with the sequence Thr-Pro-Thr-Ser, one hexapeptide with the sequence Pro-Thr-Thr-Thr-Pro-Ser and one heptapeptide with the sequence Ala-Pro-Thr-Thr-Ser-Gly-Ser. Galactosamine was present in several of these fragments and the hexapeptide and heptapeptide both contained sufficient of this sugar for all the serine and threonine residues to be glycosylated. The following five tetrapeptides sequences were deduced from mixtures of two peptides which differed by the amino-acid substituent at only one position in the chain; Pro-Thr-Ala-ser, Thr-Thr Pro-Ser, thr-Thr-Ala-Ser, Ser-Pro-Thr-Ser, and Ser-Ala-Thr-Ser. A tripeptide mixture which differed only in the nature of the third amino acid yielded the following three tripeptide sequences: Pro-Thr-Ser, Pro-thr-Ala. These peptides do not indicate a clearly defined amino-acid sequence surrounding the glycosylated hydroxy amino acids but establish that in the human blood-group-specific glycoproteins there is close packing of the threonine in the region of the peptide moiety carrying the carbohydrate chains. [ABSTRACT FROM AUTHOR]

Peptide studies on two acidic proteins, A1 and A2, from 50-S ribosomes of Escherichia coli indicate a closely related primary structure. 1. Amino acid compositions of the tryptic peptides of the two proteins are identical, with exception of the amino terminal peptide. 2. The N-terminal amino acid sequence of A2-protein is Ser-Ile-Thr-Lys, while that of A1-protein is N-acetyl-Ser-Ile-Thr-Lys. 3. The estimated number of 120 amino acid residues in each polypeptide chain is consistent with the physical molecular weight estimate. 4. In 50% of the polypeptide chains, in both A1- or A2-protein, a specific lysine residue is replaced by ε-N-monomethyl-lysine. [ABSTRACT FROM AUTHOR]

Hexameric hemocyanin from a spiny lobster. Panulirus japonicus, comprises three major subunits (Ib, II and lid end one minor subunit (Ia), as reported in the preceding paper in this journal. It has previously been shown that the O2 equilibria of Panulirus hemocyanin can be described by a concerted model extended to three affinity states [Makino, N. (1986) Fur. J. Biochem. 154, 49–55]. In this study the equilibrium binding of O2 to the reassociated sebunits (Ib, II and III) was examined at various pH in the presence or absence of Ca2+ in order to test the applicability of the three-state model to the homogeneous hexamers. The hexameric structure of the reassembled subunits was less stable than that of the native protein under the conditions examined. The model could be fitted to the O2-binding isotherms of the homohexamers composed of the subunits II or III, if the molecular dissociation of the protein was taken into account. It was postulated that the monomeric hemocyanin has the same ligand affinity as that of the hexamer in the intermediate-affinity state (S). The fitting of the model to the O2 binding of the subunit I was unsuccessful mainly because of the low cooperativity of the assembled subunits. [ABSTRACT FROM AUTHOR]

Two c-type cytochromes were isolated from cells of the gram-negative bacterium Aquaspirillum itersonii grown under low aeration in the presence of nitrate. The major component, cytochrome c-550, was equated with the (single) c-type cytochrome previously reported to be present in this organism [Clark-Walker, G. D. & Lascelles, J. (1970) Arch. Biochem. Biophys. 136, 153-159], although a significantly higher molecular mass was apparent in the present work. The complete amino acid sequence of this cytochrome is reported in the accompanying paper. A second soluble c-type cytochrome, designated c-556, was also isolated. The molecular mass, isoelectric point, spectrum, midpoint oxidation reduction potential and amino acid composition of this monoheam cytochrome are reported. The possible relationship of this cytochrome to other cytochromes c-556 is discussed. [ABSTRACT FROM AUTHOR]

The complete amino acid sequence of turnip peroxidase TP 7, the principal isoperoxidase during winter in turnip, Brassica napus L., variety Blanc dur d'hiver, has been determined by sequence analysis of cyanogen bromide fragments and of tryptic peptides. The turnip peroxidase TP 7 enzyme is composed of 296 amino acids, one hemin group and one neutral carbohydrate side chain attached through asparagine. The molecular weight of the polypeptide part is 31 060, and including heroin and carbohydrate the molecular weight of the native enzyme is close to 33 400. The isoelectric point of turnip peroxidase TP 7 is 11.6. Comparison of turnip peroxidase TP 7 and horseradish peroxidase HRP C shows that they contain four similarly located disulfide bridges and have pyrrolidone carboxylyl N termini. Their common evolutionary origin is distant as their amino acid sequences are only 49% identical. Furthermore, turnip peroxidase TP 7 differs significantly from three other isoperoxidases of turnip root, turnip peroxidases TP 1, TP 2 and TP 3, and from horseradish peroxidase HRP C in its physico-chemical and enzymatic properties, and its pronounced season-dependent appearance. All these differences of turnip peroxidase TP 7 and of the others suggest they serve separate biological functions. [ABSTRACT FROM AUTHOR]

Methods which use the scintillator PPO to record film images of ³H in chromatograms and polyacrylamide gels (fluorography) have been described elsewhere. This paper demonstrates that pre-exposure of the film to a brief flash of light greatly increases the sensitivity of fluorography. Pre-exposure also permits quantitative interpretation of the film image, because it corrects the non-linear relationship between radioactivity of the sample and absorbance of the film image. Therefore the distribution of radioactivity in the sample is accurately represented by microdensitometry of the image obtained on pre-exposed film. Using pre-exposed film 300 dis. ³H/min or 30 dis. 14C/min can be detected in a band in a gel in a 24-h exposure. The Appendix describes revisions and extensions of existing fluorographic procedures, including application to agarose gels and a rapid procedure for recovering PPO for re-use. [ABSTRACT FROM AUTHOR]

Semliki-Forest-virus 42-S virion RNA is translated in a mouse ascites cell-free system. One of the major polypeptides made in vitro has been identified as Semliki-Forest-virus capsid protein by co-electrophoresis on dodecylsulfate -- polyacrylamide gels and by tryptic peptide analysis on paper. Another major product of the cell-free system has been characterised as envelope protein by peptide analysis but it does not co-electrophorese on gels with envelope proteins from purified virions. It is possible that the nascent, non-glycosylated polypeptide is partially degraded by proleolytic enzymes present in the cell-free extract, or alternatively that fragments of envelope protein are produced by premature termination on the viral RNA. Several other polypeptides are synthesised in the cell-free system in variable amounts but these have not been identified. The genetic content and the mechanism of translation of Semliki-Forest-virus 42-S RNA are discussed. [ABSTRACT FROM AUTHOR]

With the help of an automatic protein sequenator, we have verified a prediction made by us in a recent paper, namely that residue 16 in Candida krusei cytochrome c is a glutamine and not a glutamic acid. Neurospora crassa cytochrome c now remains the only mitochondrial cytochrome c which apparently does not have a glutamine at this position. In the course of this investigation, from two strains of Candida krusei we isolated two cytochromes which differ in amino acid composition. One of them seems to correspond to that described in the literature. Two of the differences have been located. [ABSTRACT FROM AUTHOR]

In this paper we address the problem of extracting features relevant for predicting protein–protein interaction sites from the three-dimensional structures of protein complexes. Our approach is based on information about evolutionary conservation and surface disposition. We implement a neural network based system, which uses a cross validation procedure and allows the correct detection of 73% of the residues involved in protein interactions in a selected database comprising 226 heterodimers. Our analysis confirms that the chemico-physical properties of interacting surfaces are difficult to distinguish from those of the whole protein surface. However neural networks trained with a reduced representation of the interacting patch and sequence profile are sufficient to generalize over the different features of the contact patches and to predict whether a residue in the protein surface is or is not in contact. By using a blind test, we report the prediction of the surface interacting sites of three structural components of the Dnak molecular chaperone system, and find close agreement with previously published experimental results. We propose that the predictor can significantly complement results from structural and functional proteomics. [ABSTRACT FROM AUTHOR]

This paper presents the first purification of the mitochondrial branched‐chain amino acid aminotransferase (BCATm) from sheep placenta. It is a homodimer with an apparent subunit molecular mass of 41 kDa. The enzyme differs from those of the rat and human as it appears to form at least one intermolecular disulfide bond. The sheep BCATm cDNA (1.4 kb) encodes a mature polypeptide of 366 amino acids with a calculated molecular mass of 41 329 Da and a partial mitochondrial targeting sequence of seven amino acids. The sheep BCATm sequence shares higher identity with other mammalian BCATm isoenzymes (82–85%) than with the cytosolic isoenzymes (60%). By Northern blot analysis, a message of 1.7 kb was detected in sheep placenta and skeletal muscle. Measurements of BCAT activity, mRNA and BCATm protein in sheep placenta and skeletal muscle revealed that BCATm is the sole BCAT isoenzyme expressed in placenta, whereas it contributes 57 and 71% of the BCAT activity in tensor fascia latae and masseter muscles from weaned lambs respectively. Skeletal muscle, the main site of branched‐chain amino acid transamination, exhibits significantly lower BCAT activity in sheep than in rat. Our results suggest that the low BCATm mRNA level probably accounts for the low BCAT activity in sheep skeletal muscle, and that the metabolic scheme for branched‐chain amino acid catabolism is specific to each species. [ABSTRACT FROM AUTHOR]

The α2macroglobulin-receptor-associated protein (RAP) binds to the α2macroglobulin receptor/low-density lipoprotein receptor-related protein (α2MR/LRP), a multi-functional cell surface receptor known to bind and internalize several macromolecular ligands. RAP has been shown to inhibit binding of all known α2MR/LRP ligands. Mutational studies have implicated distinct parts of RAP as specifically in- volved in inhibition of binding of a multitude of ligands. In the present paper we provide experimental evidence allowing assignment of elements of triplicate internal sequence similarity in RAP, noted previously [Warshawsky, L., Bu, C. & Schwartz, A. L. (1995) Sites within the 39-kDa protein important for regulating ligand binding to the low-density lipoprotein receptor-related protein, Biochemistry 34, 3404-3415], to three structural domains, 1, 2 and 3, comprising residues 18-112, 113-218 and 219-323 of RAP, respectively. Structural analysis by 1H-NMR spectroscopy shows that domains 1 and 2 as separate domains have similar secondary structures, consisting almost exclusively of α-helices, whereas domain 3 as a separate domain appears only to he marginally stable. Ligand competition titration of recombinant RAP domains 1, 2 and 3 and double domains 1+2 and 2+3 against 125I-RAP and 125I-α2M* (methylamine-activated α22M) for binding to α2MR/LRP demonstrated (a) that functional integrity in single domains is largely preserved, and (b) that important determinants for the inhibition of test ligands reside in the C-terminal regions of domains 1 and 3. [ABSTRACT FROM AUTHOR]

Investigates the different susceptibility of protein kinase to staurosporine inhibition. Kinetic studies; Molecular bases for the resistance of protein kinase CK2.

A protein is usually classified into one of the following four structural classes: all α, all β, (a + B) and α/β. In this paper, based on the maximum correlation-coefficient principle, a new formulation is proposed for predicting the structural class of a protein according to its amino acid composition. Calculations have been made for a development set of proteins from which the amino acid compositions for the standard structural classes were derived, and an independent set of proteins which are outside the development set. The former can test the self consistency of a method and the latter can test its extrapolating effectiveness. In both cases, the results showed that the new method gave a considerably higher rate of correct prediction than any of the previous methods, implying that a significant improvement has been achieved by implementing the maximum-correlation-coefficient principle in the new method. [ABSTRACT FROM AUTHOR]

In this paper we report the development of the immunologically detected uncoupling protein (UCP) in brown adipose tissue during the perinatal period in the rat and its relationship with its functional activity expressed in terms of GDP-binding capacity, GDP-sensitive permeabilities and GDP-sensitive respiration. Immunologically detected UCP increased during the last 2 days of foetal life (under euthermic conditions) and after birth (after postnatal hypothermia) during the early suckling period, reaching its maximum value on day 10 after birth. This increase in UCP is accompanied by parallel increases in the GDP-binding capacity, GDP-sensitive permeabilities to protons and chloride ions and GDP-inhibitable respiration. During the suckling—weaning transition, there was a regression of the parameters related to the functional activity of the UCP (GDP-binding capacity and nucleotide-sensitive permeabilities and respiration) without changes in the immunologically detected UCP. These results suggest that the involution of this tissue in the rat commences in this period; the first parameters affected are those related to the functional activity of the UCP while the UCP is still present in its highest level. This seems to support the idea that, in this period of development of the rat, the UCP may exist in the brown fat mitochondria in a functional (unmasked) form and a non-functional (masked) form. [ABSTRACT FROM AUTHOR]

A pool of neutral carbohydrate chains was prepared from respiratory mucins of a patient suffering from bronchiectasis. Fractionation by HPLC led to 35 smaller-size oligosaccharide-alditols; the structure of 16 oligosaccharide-alditols with core type 1 or type 2 has been established (Klein, A., Lamblin. G., Lhermitte, M., Roussel, P., Breg, J., Van Halbeek, H. & Vliegenthart, J. F. G., preceding paper in this journal).In this second part, we identified 19 ligosaccharide-alditols possessing core types 3 and 4. Nine of the structures (1, 5, 9, 6, 10b, 13, 19, 15b and 18.1) have been described previously to be present in cystic fibrosis mucins [Lamblin, G., Boersma, A., Lhermitte, M., Roussel, P., Mutsaers, J. G. H. M., Van Halbeek, H. & Vliegenthart, J. F. G. (1984) Eur. J. Biochem. 143, 227-234]. The remaining ten are new structures isolated from bronchial mucins; they are all extensions of the above-mentioned nine oligosaccharides. These compounds are octasaccharide-alditols containing the Y determinant together with the H determinant of either backbone-type 1 or 2, and partial structures thereof:... and 8c, which is lib without Fucα(1 → 3), ...and 17b, which is 24 without Fuc and Gal in the 3-branch, and 22c, which is 24 without Fucα(1 → 3) ...and 21b, which is 23c without Fucα(1 → 3), 18.2, which is 23c without any Fuc in the 6-branch, and 22b, which is 23c without Fuc in the 3-branch. [ABSTRACT FROM AUTHOR]

A theory is proposed to explain the physical bases of the ionic control of the activity of an enzyme system, located on a plant cell surface, and probably involved in cell-wall synthesis and extension. The model, which is based on various previously published experimental results, involves several assumptions: a cell-wall pectin methyl esterase de-esterifies pectins and thus creates the fixed negative charges of the cell wall; various enzymes incorporate uncharged carbohydrates in cell-wall material; cell-wall extension implies the sliding of cellulose microfibrils; the enzymes responsible for carbohydrate incorporation are activated by protons in the pH range 4-8 and have very similar pH dependencies: the cell-wall pectin methyl esterase is inhibited by protons in the same pH range. The mathematical derivation of this model, written in the form of a hypercycle, indicates that it is equivalent to a set of two antagonistic enzyme reactions: an enzyme reaction conditioned by pectin methyl esterase which results in the increase of fixed charge density of the cell wall; a number of 'growth enzymes', which produce extension and building up of the cell wall and therefore a decrease of charge density. The mathematical study of this model shows it may display a very high co-operativity of its response to slight changes of pH. This co-operativity means that the cell wall charge density may dramatically increase or decrease, within a very narrow pH range. The steep response of this system appears to be the direct consequence of different pH sensitivities of pectin methyl esterase and of the other cell-wall enzymes involved in cell growth. Calcium, which tightly binds to the cell wall, may diminish or even suppress this abrupt charge transition. This model suggests a novel theory of the ionic control of cell-wall expansion. The very basis of this theory is the existence of an electrostatic potential difference, Δψ, between the inside and the outside of the cell wall. When this Δψ value is large, the local proton concentration is high. Therefore the enzymes involved in cell wall extension and building up are active, but pectin methyl esterase is not. Therefore, the cell wall extends and the charge density decreases. The Δψ value then declines, as well as the local proton concentration. Under these conditions, the pectin methyl esterase becomes activated, whereas the 'growth enzymes' are not. This activation of pectin methyl esterase restores the initial, or an even higher, clectrostatic potential difference, which in turn results in a decrease of local pH. In the following papers the basic ideas of this model are subjected to experimental control. [ABSTRACT FROM AUTHOR]

Isolated chloroplasts of Acetabularia incorporate radioactive amino acids into more than 30 polypeptides in the light, including the apoprotein of the P700-chlorophyll a protein complex, the reaction centre core of photo- system I [Biochim. Biophys. Acta, 609, 107–120 (1980)]. In this paper it is shown that the apoproteins of the two minor chlorophyll a complexes, thought to be part of photosystem II reaction centre core, are also syn- thesized by isolated chloroplasts. Furthermore, they are integrated correctly into the thylakoid membrane in the absence of any cytoplasmic contribution, such that they can be isolated as chlorophyll-protein complexes indistinguishable from those already in the membrane. In contrast, the minor chlorophyll a + b complex ‘CP 29’ [Camm, E. L. and Green, B. R. (1980) Plant Physiol. 66, 428–432] and its dimers are not synthesized by isolated chloroplasts. In this they resemble the other chlorophyll a + b complex, the light-harvesting complex (LHC). It may be significant that the LHC, which is not essential for photosynthetic activity, is under nuclear control, while the reaction centre polypeptides, cytochrome b559, and cytochrome f, are synthesized on chloroplast ribosomes. [ABSTRACT FROM AUTHOR]

The brain specific S-100 protein is a mixture of two components S-100a and S-100 b with a subunit composition αβ or β2 respectively. The amino acid sequence of the β subunit has been previously determined. This paper presents the sequence of the α subunit in the S-100a protein. The α subunit consists of 93 amino-acid residues and has a relative molecular mass of 10400. The sequence shows extensive homology (58 %) with that of the β-subunit and shares an apparent calcium binding site in the C-terminal half of the molecule, suggesting a close evolutionary relationship between these subunits. [ABSTRACT FROM AUTHOR]

The complete primary structure of the constant part of the μ-chain-disease protein, BOT, was established. It includes the whole CH2, CH3 and CH4 domains. Two amino acid changes were found, at positions 309 (Ser ↵ Gly) and 333 (Val ↵ Gly) (GAL numbering). In two additional monoclonal n chains (SCO and CO), the same positions showed an amino acid variability. From these data it may be concluded that four types of μ chains exist in the human: (1) GAL type with Ser-309 and Val-333; (2) OU type with Gly-309 and Val-333; (3) SCO type with Ser-309 and Gly-333; (4) BOT/CO type with Gly-309 and Gly-333. The meaning of this molecular polymorphism is discussed. [ABSTRACT FROM AUTHOR]

This paper considers how apolipoprotein CII from human plasma lipoproteins and T1 and T2 proteins from egg yolk Iipoproteins stimulate the activity oflipoprotein lipase. These activator proteins stabilized the enzyme much more effectively than a thousandfold higher concentration of albumin did, indicating a direct interaction with the enzyme. The effects of the activators were seen also at 1 M NaCl. Thus, forces other than electrostatic are implicated. Centrifugation experiments showed that 125I-labeled lipase bound equally well to the emulsion droplets in the absence of activator protein as in its presence. This was true even under conditions when the activator caused a severalfold increase in the rate of hydrolysis. Thus, the activator makes enzyme at the interface more effective in hydrolysis. By optimizing the conditions it was possible to obtain almost as high rates of triglyceride hydrolysis in the absence as in the presence of activator. Thus, the main effect of the activator protein is probably not on a rate-limiting chemical step. Under most conditions, the rate of hydrolysis was much below optimal and activator increased it. This was always the case with phosphatidyl-choline/triglyceride emulsions, where the activator enhanced hydrolysis of both lipids. Other experiments showed that the activator enhanced triglyceride hydrolysis in the absence of phospholipids and phospholipid hydrolysis in the absence of triglycerides. It is suggested that interaction with activator orientates the enzyme and/or the lipid substrate for effective hydrolysis at the surface of lipoproteins/model substrates. [ABSTRACT FROM AUTHOR]

Glycophorin can be incorporated in lipid vesicles without the use of detergents [MacDonald, R. J. and MacDonald, R. C. (1975) J. Biol. Chem. 250, 9206–9214]. In this paper it has been shown that the protein spans the lipid bilayer in these vesicles. In addition it has been shown that the protein is orientated in an asymmetric way with 75–80% of its sugar residues directed to the outside of the vesicles. Freeze-fracturing electron microscopy suggests that the protein is present in a slightly ag- gregated form. Sonication of these recombinants leads to the formation of small glycophorin-containing vesicles, in which almost all sugar residues are directed to the outside. [ABSTRACT FROM AUTHOR]

A parvalbumin-like protein was purified to homogeneity from chicken leg muscle. 40 mg of protein was isolated from 1 kg muscle which is 20% of the amount obtained for parvalbumin isolated from rabbit muscle by a similar procedure. The protein contained 1-2 residues of tyrosine and 1 residue of tryptophan, three arginine residues and a rather small number (4-5) of phenylalanine residues. The differences in the aromatic amino acid content account for the less distinct vibronic structure in the ultraviolet region below 275 nm compared to most other parvalbumins. The protein contained <0.04 residue of γ-carboxyglutamic acid per molecule. However, the molecular weight of 12 000, high content of glutamic acid and aspartic acid, the low content of proline and histidine, and the presence of two calcium atoms per molecule were properties common to parvalbumins. Immunprecipitation experiments in agar using anti-chicken parvalbumin-like protein indicated that an immunologically indistinguishable form of this protein is present in heart, white and red back muscles, breast, stomach, brain, pancreas, small intestine and kidney, but not in spleen, liver and blood. The distribution is therefore as ubiquitous as G-actin or β-actinin. Parvalbumin-like protein was found to accumulate in differentiating primary muscle cell cultures after 120 h. This approximates to the time on which myosin and actin start to accumulate, proteins necessary for myofibril assembly. Experiments to investigate the localization of parvalbumin-like protein, using the indirect immunofluorescence technique showed regular cross-striation patterns within the I band of isolated myofibrils. The results suggest that parvalbumin-like protein may interact with one or more of the protein components of the thin filament thus being involved in the contractile process. This component could be β-actinin which copurified with parvalbumin-like protein through the DEAE-cellulose step (see following paper). [ABSTRACT FROM AUTHOR]

The primary structure of the major parvalbumin (pI = 4.45) from the cartilaginous fish Raja clavata has been determined. The amino acid sequence was deduced by the analysis of peptides derived from tryptic digestion of the oxidized protein. These peptides were aligned by comparison with (a) overlapping peptides produced by limited tryptic and chymotryptic digestion, and (b) by comparison with the known structures of other fish parvalbumins. The molecular evolution of thornback ray parvalbumin is briefly discussed. [ABSTRACT FROM AUTHOR]

Sheep-serum transferrin shows marked polymorphism and more than 20 alleles have been identified although only 4 or 5 of these have a frequency higher than 1%. Each of the alleles has two bands in starch-gel electrophoresis, corresponding to a major and a minor fraction. This paper describes the isolation and partial characterisation of the two fractions from the transferrin of sheep homozygous for Tf B. The purification consisted of: (a) precipitation by ammonium sulphate, (b) chromatography on CM-cellulose and, finally (c) chromatography on DEAE-Sephadex. The purification procedure had no effect on the electrophoretic mobility of the two fractions and they appeared homogeneous by starch-gel and polyamide-gel electrophoresis and by immuno-electrophoresis. The amino-acid compositions of both fractions were very similar and the sequences of the first eight amino-acid residues: Ser-Pro-Glu-Lys-Thr-Val-Arg-Trp-were identical for both bands. These results, and the fact that the two fractions are found in all genetic variants and always have the same relative mobility strongly suggest that the differences do not lie in the polypeptide chain. From the results of hydrolysis by neuraminidase and assay of sialic acid, the major and minor fractions most probably contain two and three sialic acid residues (exclusively N-acetylneuraminic acid) respectively, thus explaining the different electrophoretic mobilities. The sialic-acid content has been calculated on the basis of a molecular weight of 77500 as determined both by low-speed equilibrium ultracentrifugation and by Archibald's method. The following physico-chemical constants have been established: absorption coefficient at 280nm, A1 mg/cm³1 cm = 1.25 ± 0.01 cm² · mg-1; partial specific volume, &vmacr; = 0.734 cm³ · g-1; sedimentation coefficient s20.wo = 5.15 S; refractive-index increment, dn/dc = 0.173 cm³ · g-1. [ABSTRACT FROM AUTHOR]

In this paper the mode of action of IF-1 in 40-S initiation complex formation was studied with MS 2 RNA as messenger. Using initiation factors IF-2 and IF-3 labeled in vitro it appeared that IF-I did not influence the binding of these factors in the absence of fMet-tRNA. However, in the presence of fMet-tRNA it was found that the enhancement of the fMet-tRNA binding by IF-1 was accompanied with an equimolar increase in binding of IF-2. Moreover, it appeared that also in absence of IF-I, fMet-tRNA binding is coupled with an equimolar enhancement of the IF-2 binding, which suggests the existence of a preribosomal complex between IF-2 and fMet-tRNA. The apparent K,, values for both the binding of fMet-tRNA and IF-2 to 30-S subunits were determined and appeared to be equal, which makes a functioning of such a preribosomal complex in protein initiation very likely. The participation of GTP in this complex will be discussed. Functions of IF-I in dissociation and recycling of IF-2, described by others, and the stimulation on the 30-S subunit level might well be explained as pleiotropic effects of one basic action of IF-1, i.e. a conformational change of 30-S subunits. [ABSTRACT FROM AUTHOR]

The primary structure of protein S8 from the 30-S ribosomal subunit of Escherichia coil was determined mainly by automatic Edman degradation using a modified Beckman protein sequenator and the solid-phase sequenator of Laursen. The complete sequence, containing 109 amino acids, was derived by analysing peptides from tryptic, chymotryptic, thermolysin, staphylococcal protease and cyanogen bromide digestion of the protein. The amino acid composition was found to be (aspartic acid)6, (asparagine)3, (threonine)5, (serine)5, (glutamic acid)7, (glutamine)6, (proline)5, (glycine)6, (alanine)11, (valine)9, (methionine)4, (isoleucine)7, (leucine)9, (tyrosine)3, (phenylalanine)3, (lysine)11, (arginine)8, (cysteine)1. S8 is a basic protein and binds to the 16-S RNA; knowledge of its sequence is necessary for a detailed study of its interaction with the ribosomal RNA. [ABSTRACT FROM AUTHOR]

A procedure is described for the simultaneous separation of all the peptides (other than those present in trace amounts) contained in the venom of the common European honey bee (Apis mellifera). Three new peptides have been isolated: one is a fragment of the known peptide melittin. Some properties of the peptides are described. [ABSTRACT FROM AUTHOR]

We have further characterized the polynucleotide ligase purified from cultures of the human heteroploid line EUE [Spadari, Ciarrocchi and Falaschi, Eur. J. Biochem. 22, 75 (1971)]. The 350-fold purified enzyme gives positive response with four different assays; the rates with the different substrates are different, but the variations are identical to those observed with the purified ligase induced by T4-phage. The enzyme can reconstitute the transforming activity of Bacillus subtilis DNA inactivated by pancreatic DNAase. The human cell enzyme is unable to use a hybrid substrate where an interrupted polydeoxynucleotide is annealed to a polyribonucleotide, whereas in the same conditions the T4 enzyme gives appreciable activity. The partial dependence of the purified enzyme on proteins present in a boiled crude extract, reported in the previous paper, seems due to an aspecific protective effect of the soluble proteins of cell extracts. The enzyme does not show any appreciable sequence specificity in the phosphodiester bond it can form. The purified enzyme can be fractionated into two molecular forms, one having a molecular weight of 190000, the other 95000; fresh extracts of EUE cells contain almost exclusively the high molecular-weight form; ageing of the extract or purification lead to the appearance of the second peak, without variations in the total activity. This could correspond to the conversion of a dimer structure into a monomer. The “dimer” has a pH optimum close to 8.1, whereas the “monomer” has its optimum at 7.5; this explains the bimodal pH curve previously described in the purified enzyme. [ABSTRACT FROM AUTHOR]

Binding of tryptophan, tryptamine and ATP to tryptophanyl-tRNA synthetase was studied by equilibrium dialysis experiments. There arc two binding sites per mole of enzyme both for tryptophan and for tryptamine. Ks for tryptophan is 0.95 μM and for tryptamine is 1.8 μM. In the case of ATP no binding could he measured over the range of concentrations examined. The Scatchard plots of tryptophan and tryptamine binding do not shown any cooperativity between the subunits. Dissociation of the dimeric enzyme was studied at very low protein concentration. Upon dilution of the enzyme both the [32P]PPi-ATP isotope exchange activity and the tRNA charging activity are lost simultaneously. Kinetic evidence demonstrates that the inactivation is primarily due to the dissociation of the active direct into inactive monomers. The dissociation is strongly promoted by alkaline pH and is inhibited by the presence of tryptophan or tryptophanyladenylate but not by that of tRNA. The dissociation constant of the dimer-monomer equilibrium at pH 8.5 is 15 nM at 25 °C. The dissociation form is more susceptible to denaturation than the dimeric species. The results reported in the paper suggest that even if there are not binding interactions between the subunits, the active conformation of the enzyme depends on their associated state. [ABSTRACT FROM AUTHOR]

The αA2 chain of bovine α-crystallin was fragmented by means of cyanogen bromide treatment and by tryptic, chymotryptic and thermolytic digestions. Twenty tryptic peptides were obtained from the S-aminoethylated αA2 chain, accounting together for the complete sequence. The direct Edman degradation and the dansyl-Edman technique were used to determine the sequences of the tryptic peptides. The order of the tryptic peptides was deduced from overlapping peptides obtained by cyanogen bromide treatment, tryptic digestion of the maleylated chain and chymotryptic and thermolytic digestion of the S-aminoethylated chain. The sequence of the αA2 chain comprises 173 residues and corresponds to a molecular weight of 19832. [ABSTRACT FROM AUTHOR]

The isolation of oligosaccharides produced by the action of phage-induced enzymes on the bacterial exopolysaccharide colanic acid is reported. The products from colanic acid produced by different strains of Escherichia coli and Salmonella typhimurium were hexasaccharides which differed in their acyl substituents. All contained fucose, glucose, galactose and glucuronic acid in the molar ratio 2:1:2:1. They appeared to have the same carbohydrate structure as that postulated for the repeating unit of colanic acid. All the hexasaccharides contained acetate. Pyruvate was also found in most preparations. The phage-induced enzymes only converted 30-35 % of the polymer to oligosaccharides. They also hydrolysed de-acetylated polysaccharide but, were inactive against carboxyl-reduced material and against a number of other bacterial exopolysaccharides. [ABSTRACT FROM AUTHOR]

The analytical active-enzyme-centrifugation method permits the determination of the hydrodynamic properties of enzymes while they are fully active on their substrates: the centrifugation can be performed at the very dilute concentrations used in kinetic studies (&aysmp; 1 µg/ml): furthermore a purified enzyme preparation is not necessary and in many cases a crude extract is sufficient. This method is beginning to be used in several laboratories even though few of its experimental aspects have been published. This paper reports the details for carrying out, an activeenzyme-centrifugation run, discusses the experimental conditions anti details with artifacts and ways, which are easy, to overcome them. [ABSTRACT FROM AUTHOR]

Semihemoglobins α and β are derivatives of human hemoglobin which possess the usual chain composition (α2β2 or αβ) but which carry the heine prosthetic group on only one kind of chain (α or β), the complementary chain (β° or α°) being heme-free. In this paper, new results on two compounds of this type, namely, semiHbαD (αβ°) and semilHbβ (α°β) have been presented; existing data on other earlier preparations have also been reviewed. Methods for preparation of semiHbαD and semiHbβ have been described-. Some of their properties including optical absorption, circular dichroic spectra, oxygen equilibrium and kinetics of carbon monoxide binding have been measured; these properties are then compared with those possessed by intact hemoglobin (β2β2), isolated chains (α,β) and the berne-free apoprotein (α°β°). 1. The contribution of the heine moiety to the optical absorption and circular dichroic spectra of the semihemoglobins are found to be intermediate between those of hemoglobin and those of the corresponding isolated chain. This result indicates that the effect of interaction between the unlike chains of these molecules are transmitted as far as the heine prosthetic group. The structure of the heme-free chain is also modified as the result of this interaction as revealed by titration of sulfhydryl groups, although circular dichroic spectra in the far ultraviolet are not conclusive. 2. The oxygen affinity and the Bohr effect of the semihemoglobins are again intermediate between those of hemoglobin and isolated chain, the semiHbβ being closer to hemoglobin than semiHbαD. The same is true concerning the velocity of carbon monoxide binding. 3. The kinetics of fixation of carbon monoxide to semihemoglobins is strongly biphasic. It is suggested that this property could reflect the existence of semihemoglobins as two types of filmers (αβ) but differing among themselves on account of different contact regions brought into play. (α1β1 and α1β1 in the nomenclature of Perutz.) 4. The significance of the fact that semiHbα is obtained in multiple forms is considered. 5. The ensemble of the results are discussed in the context of chain-chain interactions. [ABSTRACT FROM AUTHOR]

The Cbl-1 mutant of Bacillus subtilis 168 WT is the result of a spontaneous mutation presenting a pleiotropic character. In this paper, the results of the morphological and chemical analyses of the cell walls isolated from both strains are compared. When either the whole cells or the isolated cell walls of both strains were fixed with osmium tetroxide, the examination by electron microscopy showed extensive morphological differences in structure due to the partial destruction of the mutant cell walls. The chemical investigations performed on the isolated cell walls revealed surprising differences in chemical structure. In the cell walls of the mutant, teichoic acid was completely missing and was replaced by protein. A major protein-containing fraction accounting for about 50% of the material solubilized by autolysis of the mutant cell walls was isolated. Polyacrylamide gel electrophoresis showed that this fraction was composed mainly of a single protein of high molecular weight When denatured by sodium dodecyl sulfate treatment in the presence of β-mercaptoethanol, the protein yielded by gel electrophoresis two major bands with molecular weights of approximately 105000 and 155000, respectively. The molecular weight of the Cbl-1 cell wall protein determined by high speed sedimentation equilibrium was 255 600. [ABSTRACT FROM AUTHOR]

The study of the structure of the peptides arising from native, oxidized or reduced and maleinized bovine growth hormone on incubation with trypsin, ehymotrypsin and pepsin is reported. The data obtained permitted the assembly of a unique sequence of amino acids for the poly-peptide chain of the protein. Various corrections and one addition to the sequence previously communicated are made. [ABSTRACT FROM AUTHOR]

A protein is generally classified into one of the following four structural classes: all α, all β, α+β and α/β. In this paper, based on the weighting to the 20 constituent amino acids, a new method is proposed for predicting the structural class of a protein according to its amino acid composition. The 20 weighting parameters, which reflect the different properties of the 20 constituent amino acids, have been obtained from a training set of proteins through the linear-programming approach. The rate of correct prediction for a training set of proteins by means of the new method was 100%, whereas the highest rate of previous methods was 82.8 %. Furthermore, the results showed that the more numerous training proteins, the more effective the new method. [ABSTRACT FROM AUTHOR]

In mouse spermatozoa, DNA is compacted by two protamines mP1 and mP2. Protamine mP2 (63 residues) is synthesized in spermatid nuclei as a precursor pmP2 (106 residues) which is subsequently processed at the end of spermiogenesis [Yelick, P. C., Balhorn, R., Johnson, P. A., Corzett, M., Mazrimas, J. A., Kleene, K. C. & Hecht, N. B. (1987) Mol. Cell. Biol. 7, 2173-2179]. Six proteins, three of which were described earlier [Chauvière, M., Martinage, A., Debarle, M., Alimi. E., Sautière, P. & Chevaillier, Ph. (1991) C. R. Acad. Sci. 313, 107-112], have molecular and electrophoretic properties similar to those of pmP2. They were isolated from purified testis nuclei and characterized by amino acid composition, N-terminal sequence and peptide mapping. From the amino acid compositions, it appears that all six proteins are rich in arginine, cysteine and histidine and are closely related to pmP2 and mP2. The N-terminal sequence of each protein overlaps a distinct region of the N-terminal part of pmP2. The C-terminal part of protamine mP2 starting at arginine 15 is common to all proteins as assessed by amino acid compositions and peptide maps, ll these structural data demonstrate that the six isolated proteins are products of pmP2 precursor processing. The six intermediate proteins pmP2/5, pmP2/11, pmP2/16, pmP2/20, pmP2/26 and pmP2/ 32 which contain 102, 96, 91, 87, 81 and 75 residues, respectively, are generated from the pmP2 precursor after N-terminal excision of 4, 10, 15, 19, 25 and 31 residues, respectively. The C-terminal sequence of protamine mP2 is strictly identical to that of its precursor; therefore, no maturation occurs in this part of the molecule. At the present time, the proteolytic pathway involved in the amino-terminal processing leading to the mature form of the protamine mP2 (63 residues) has not been elucidated. However, the different representation of six intermediates in the testis suggests that some stages of processing are faster than others or that some cleavage sites are preferred. The proteins described in this paper could result either from stepwise excision of N-terminal residues or from nonsequential cleavages. [ABSTRACT FROM AUTHOR]