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Membranes from human placenta contain proteins which inhibit the activity of phospholipases A2 by binding to phospholipid thus impeding substrate availability. We used unilamellar mixed liposomes and a partially purified cytosolic phospholipase A2 from placenta for characterizing this substrate-depleting activity. A major portion of these inhibitory proteins was released by extracting washed membranes with a Ca+(+)-chelator. Biochemical fractionation and systematic analysis resulted in the unequivocal identification of a series of annexin proteins. We describe a straightforward procedure which allows to obtain 8 annexins from placenta either in pure form or as a mixture of two annexins. One of them was obtained in two forms which had the same molecular mass of 68 kDa but differed in charge. We also present suggestive evidence for a novel annexin I-related polypeptide of Mr 45,000 which is an excellent in vitro substrate for protein kinase C. We estimate that about 2% of the total placental membrane proteins are annexins. For achieving half inhibition of phospholipase A2 activity with pure annexins, up to a 6.5-fold difference in the amounts of protein was observed when calculated on a molar basis. This suggests specificity of individual annexin species.

New membrane-preference scales are introduced for categories of membrane proteins with different functions. A statistical analysis is carried out with several scales to verify the relative accuracy in the prediction of the transmembrane segments of polytopic membrane proteins. The correlation between some of the scales most used and those calculated here provides criteria for selecting the most appropriate methods for a given type of protein. The parameters used in the evaluation of the hydropathy profiles have been carefully ascertained in order to develop a reliable methodology for hydropathy analysis. Finally, an integrated hydropathy analysis using different methods has been applied to several sequences of related proteins. The above analysis indicates that (a) microsomal cytochrome P450 contains only one hydrophobic region at the N-terminus that is consistently predicted to transverse the membrane: (b) only four of the six or seven putative transmembrane helices of cytochrome oxidase subunit III are predicted and correspond to helices I, III, V and VI of the previous nomenclature; (c) the product of the mitochondrial ATPase-6 gene (or the chloroplast ATPase-IV gene) of F0-F1-ATPase shows that helix IV is not consistently predicted to traverse the membrane, suggesting a four-helix model for this family of proteins.

The uncoupling protein found in mitochondria from thermogenic brown adipose tissue is structurally very similar to two other mitochondrial carrier proteins transporting ADP/ATP and phosphate, respectively. Similarities are also seen with the mechanism of these carriers, which are part of a family of H+/OH(-)-substrate anion co-transporters, further strengthening the evidence that the uncoupling protein has evolved from this family of mitochondrial carrier proteins.

Group B Neisseria meningitidis is thus far subdivided into 15 protein serotypes based on antigenically different major outer membrane proteins. Most serotypes have three or four major proteins in their outer membranes. Comparative structural analysis by chymotryptic 125I-peptide mapping was performed on these major proteins from the prototype strains as well as from six non-serotypable strains. The major outer membrane proteins from each of the serotypes were first separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis using the Laemmli system. Individual proteins within the gel slices were radioiodinated and digested with chymotrypsin, and then their 125I-peptides were separated by electrophoresis and chromatography on cellulose thin-layer plates. The peptide maps obtained by autoradiography were categorized into five different structural classes which correlated with the apparent molecular weights of proteins, i.e., 46 +/- 1K, 41 +/- 1K, 38 +/- 1K, 33 +/- 1K, and 28 +/- 1K. Each of the major outer membrane proteins within a strain had a distinctly different chymotryptic peptide map, indicating significant differences in the primary structure of these proteins. In contrast, outer membrane proteins of the same or very similar molecular weight from different serotype strains had similar, occasionally identical peptide maps, indicating a high degree of structural homology. The unique peptides from proteins of the same structural classes were often hydrophilic, whereas common peptides were often hydrophobic, suggesting that the serotype determinants reside within the variable hydrophilic regions of major outer membrane proteins.

Each subunit of the human transferrin receptor (TR) dimer is inserted into the ER membrane as a transmembrane polypeptide having its N-terminus in the cytoplasm. The transmembrane segment of the molecule serves both as a signal for chain translocation and as a membrane anchor. To study which structural features of this segment are required for its dual function, we have essentially replaced the transmembrane peptide with the C-terminal membrane-spanning segment of two proteins having a separate N-terminal translocation signal and with an artificial uncharged peptide. In each case the mutant TR molecules are efficiently translocated in vitro. In contrast, substitution of the transmembrane peptide of TR with a hydrophilic peptide results in no detectable translocation activity of the mutant TR. This suggests that the hydrophobic character of the transmembrane peptide of TR, rather than its actual amino acid sequence, is important for chain translocation and membrane binding.

The protein composition of membranes was studied in 17 Bdellovibrio strains by electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate. No similarity in the protein composition of membranes was found in the strains grown on cells of one and the same host. A dendrogram constructed basing on the similarity coefficients between the strains allowed to subdivide them into 3 groups according to the protein composition of their membranes. This correlated with the other phenotypic features and genotaxonomic data.

The lipopolysaccharide (LPS) and heat-modifiable outer-membrane protein (P') serotypes of 39 coded strains of group-A Neisseria meningitidis isolated from patients during seven geographically and temporally separate outbreaks of infection were determined blindly. LPS serotype discriminated between strains from different outbreaks and between strains of differing sulphadiazine sensitivity within a single outbreak. Thirty-seven strains were of three separate serotypes and no strain was of multiple serotypes. In contrast, P' serotypes did not discriminate between strains. Multiple serotypes for single strains and among strains from a single outbreak were the rule. LPS serotyping appears to be a useful epidemiological tool for distinguishing group-A strains of N. meningitidis.

Gonococcal outer-membrane protein I (PI) and PIII were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from reduced and unreduced whole-cell and outer-membrane lysates of four strains of nonpiliated (P-), transparent (O-) Neisseria gonorrhoeae. These proteins were radioiodinated and digested with alpha-chymotrypsin. The resultant 125I-peptides were then resolved by high-voltage thin-layer electrophoresis, followed by ascending thin-layer chromatography, and visualized by autoradiography. Results corroborated previous observations regarding the structural relationships of PIs having different apparent subunit molecular weights. All PIIIs had very similar apparent primary structures, regardless of the strain from which they were isolated, the source (i.e., whole cells or outer membranes), or the reduction state of the sodium dodecyl sulfate lysates. By the techniques used, it appeared that PIII is structurally similar in all of the gonococcal strains studied, even though each strain had structurally unique PIs.

Enteropathogenic Escherichia coli belonging to O groups O111 and O55 and isolated from cases of infantile diarrhea in 26 countries all over the world during 1950-1960 were examined for their outer-membrane protein (OMP) patterns; nine O:H serotypes of O111 and 11 serotypes of O55 were represented. Characteristic biotypes have earlier been described as closely associated with the most common O:H serotypes: O111:H2, O111:H12, O55:H6, and O55:H7. Different OMP patterns characterized each of the common O:H serobiotypes. The OMP patterns can be looked upon as another set of stable phenotypic characters in addition to the serotype antigens and the fermentative characters. The description of geographically widely spread, stable sero-/bio-/OMP types supports the concept that the typical enteropathogenic E coli strains also have a clonal connection.

Alkaline phosphatase (APase) has been previously described as a membrane marker correlating with B cell proliferation after stimulation by selected B cell mitogens. We have found, however, that the appearance of B cell membrane APase correlates more closely with differentiation than with proliferation. This conclusion has been drawn from the following observations: 1) APase activity appears well after peak B cell thymidine uptake, 2) mitogens which stimulate only B cell proliferation (Salmonella typhimurium mitogen) fail to induce expression of the enzyme, and 3) when proliferation of mitogen-activated B cells is inhibited, APase activity is not suppressed and may even be augmented. In addition to membrane expression, APase is also spontaneously shed into the surrounding milieu, perhaps as a result of endogenous phospholipase activity. By using a group of well-characterized inhibitors, the APase activity was shown to belong to class I (similar to the bone/liver/kidney class). Because APase always appears in differentiating but not proliferating cells, we would propose that the enzyme appearance is a late marker of B cell activation, associated with cell progression to differentiation and consequent IgM synthesis.

A high annual incidence of meningococcal meningitis and septicemia occurred in Spain from 1976 through 1980 with a peak of 19 cases per 100,000 population in 1979. Approximately 80% were caused by group B Neisseria meningitidis. Studies were undertaken to determine the distribution of groups, outer membrane protein serotypes and polyacrylamide gel electrophoresis (PAGE) types among 338 disease-associated group B isolates from six regions of Spain. The related serotypes 1, 8, and 15 accounted for 38% (129 of 338) of the isolates. Serotype 2, the major disease type in the United States, was responsible for 14% (48 of 338) of the disease in Spain and was prevalent in only one region. Forty-three percent (146 of 338) were nonserotypable. The predominant PAGE type among the nonserotypable strains was PAGE type IV (79%). These studies demonstrate the necessity of surveillance for selection of suitable serotypes to be included in protective group B meningococcal vaccines.

Several recent discoveries indicate that the ras genes, frequently activated to a transforming potential in some human tumours, belong to a large family that can be divided into three main branches: the first branch represented by the ras, ral and rap genes; the second branch, by the rho genes; and the third branch, by the rab genes. The C-terminal end of the encoded proteins always includes a cystein, which may become fatty-acylated, suggesting a sub-membrane localization. The ras superfamily proteins share four regions of high homology corresponding to the GTP binding site; however, even in these regions, significant differences are found, suggesting that the various proteins may possess slightly different biochemical properties. Recent reports show that some of these proteins play an essential role in the control of physical processes such as cell motility, membrane ruffling, endocytosis and exocytosis. Nevertheless, the characterization of the proteins directly interacting with the ras or ras-related gene-products will be required to precisely understand their function.

Six cell surface glycoproteins defined by monoclonal antibodies were selected for study on human epidermal cells. In tests on tissue sections, three of the glycoproteins [J143 (gp140/30); T43 (gp85/36); H99 (gp38)] were expressed in the basal cell layer of the epidermis, whereas the other three glycoproteins [T179 (gp140/95); T16 (gp40/50); BT15 (gp80)] were preferentially expressed in maturing keratinocytes above the basal layer. We compared synthesis of these glycoproteins in fresh epidermis and in primary epidermal short term cultures using [35S]methionine for metabolic labeling. Synthesis of J143 was 8- to 20-fold higher and synthesis of T43 was 4- to 10-fold lower in cultured cells compared with fresh epidermis. BT15, an antigen strongly expressed on terminally differentiating keratinocytes, was synthesized at 5- to 15-fold higher levels in fresh epidermis than in cultured cells. Biosynthesis levels of H99, T179, and T16 did not change in cultured epidermal cells. Based on our findings, we propose a model of surface antigenic changes that occur during keratinocyte differentiation in vivo.

Isolates of Haemophilus influenzae type b with a distinctive outer-membrane protein subtype, designated 1H, were responsible for 11 of 13 cases of invasive infection in children attending six day-care centers with secondary cases of disease, in comparison to three of 14 cases in children attending 10 centers with single cases and two centers with unrelated cases. The other 11 cases were caused by type b isolates of six different subtypes. The secondary attack rate in contacts younger than four years of age exposed to disease due to isolates of the 1H subtype was 16 per 1,000, compared with 1.6 per 1,000 after exposure to disease caused by subtypes other than 1H (P less than 0.008). Carriage rates were increased in day-care-center contacts exposed to patients with disease, and most of the carriers were colonized by isolates with subtypes homologous to those responsible for the respective index cases. Both 1H and non-1H strains were capable of colonizing contacts, but 1H strains may be more pathogenic than other subtypes.

Retina cognin (R-cognin) is an intrinsic membrane protein of vertebrate retinal cells which supports tissue-specific cell adhesion and mediates cell type-specific associations during development. As a first step in understanding how R-cognin mediates specific adhesion of retinal cell membranes, we asked if cognin bound to another cognin molecule or to a different macromolecule, a possible cognin-binding protein. To do this, we constructed an affinity column with retinal cell membrane proteins (enriched for cognin) bound to the matrix. Proteins in a detergent extract of retinal cell membranes were exposed to this matrix and those which bound specifically eluted and identified by immunoelectrophoresis. Most prominent among these was a protein with an apparent mass of 64 kDa. The binding of this material to the column was blocked by cognin antibody. To eliminate possible artifacts of molecular interactions in vitro, we sought independent confirmation that 64 kDa protein actually bound R-cognin. Using a modified retina membrane vesicle system, we asked what proteins could be photoaffinity cross-linked to cognin during vesicle aggregation. Cross-linking produced a 114 kDa complex on gels which could be resolved into a 50 kDa (cognin) and a 64 kDa band under reducing conditions. Identification of a 64 kDa protein by independent techniques suggests that cognin promotes association of embryonic chick neural retina cells by binding to this macromolecule or these molecules. Identification of a second component in the mechanism should allow elucidation of cognin's role in mediating cell-cell interactions in developing neural retina.

Over a 12-month period we obtained throat cultures from 1,448 children less than 5 years of age attending well-child clinics and identified 24 carriers of Haemophilus influenzae type b (1.7%). The outer membrane protein subtypes of the strains from the carriers were compared to the subtypes of isolates from 50 patients with Haemophilus type b disease hospitalized in St. Louis, Mo., during the same period (1981 to 1982), and the latter were compared to the subtypes of isolates from 51 patients hospitalized between 1977 and 1980. There were no significant differences in the frequencies of the five most common subtypes (1L, 1H, 2L, 2H, and 3L), comparing isolates from the carriers to those from the patients. However, 5 of the 24 throat isolates had the unusual 13L subtype compared with only 1 of the 50 invasive isolates (P = 0.02). The lower frequency of 13L strains among the invasive isolates suggests that type b isolates with this subtype may be less pathogenic than type b isolates with other subtypes. Subtype 2L strains accounted for only 2% of recent cerebrospinal fluid or blood isolates, compared with 22% of those from 1977 to 1980 (P = 0.02). Subtype 1H and 3L strains together accounted for 73%, compared with 47% of the earlier ones (P = 0.02). Thus, temporal shifts may also occur in the subtype distribution of Haemophilus type b strains causing invasive disease in a community.

Bacterial membranes have diverse functions, depending on whether they are specialized membranes or cytoplasmic membranes possessing transport, mitochondrial activities and biosynthetic functions for assembly of membranes, walls and capsules. In contrast to plasma membranes which serve as major biochemical organelles of both Gram-positive and Gram-negative bacteria, the outer membranes of the latter group confer barrier functions on the cells, providing a variety of selective channels. Although prokaryotic cells lack the array of membranous organelles characteristic of eukaryotic cells, bacteria with specific physiological and genetic capabilities form specialized membrane systems such as the bacteriorhodopsin purple membrane, chromatophore membranes of phototrophs, and forespore membranes essential to bacterial endospore formation. Unravelling the structure, function and proteins of these membranes presents a formidable biochemical, immunochemical and structural challenge.

We have investigated the nature and distribution of different populations of heparan sulfate proteoglycans (HSPGs) in several cell lines in culture. Clone 9 hepatocytes and NRK and CHO cells were biosynthetically labeled with 35SO4, and proteoglycans were isolated by DEAE-Sephacel chromatography. Heterogeneous populations of HSPGs and chondroitin/dermatan proteoglycans (CSPGs) were found in the media and cell layer extracts of all cultures. HSPGs were further purified from the media and cell layers and separated from CSPGs by ion exchange chromatography after chondroitinase ABC digestion. In all cell types, HSPGs were found both in the cell layers (20-70% of the total) as well as the medium. When the purified HSPG fractions were further separated by octyl-Sepharose chromatography, very little HSPG in the incubation media bound to the octyl-Sepharose, whereas 40-55% of that in the cell layers bound and could be eluted with 1% Triton X-100. This hydrophobic population most likely consists of membrane-intercalated HSPGs. Basement membrane-type HSPGs were identified by immunoprecipitation as a component (30-80%) of the unbound (nonhydrophobic) HSPG fraction. By immunofluorescence, basement membrane-type HSPGs were distributed in a reticular network in Clone 9 and NRK cell monolayers; by immunoelectron microscopy, these HSPGs were localized to irregular clumps of extracellular matrix located beneath and between cells. The cells did not produce a morphologically recognizable basement membrane layer under these culture conditions. When membrane-associated HSPGs were localized by immunoelectron microscopy, they were found in a continuous layer along the cell membrane of all cell types. The results demonstrate that two antigenically distinct populations of HSPG--an extracellular matrix and a membrane-intercalated population--are found at the surface of several different cultured cells lines; these populations can be distinguished from one another by differences in their distribution in the monolayers by immunocytochemistry and can be separated by hydrophobic chromatography; and basement membrane-type HSPGs are secreted and deposited in the extracellular matrix by cultured cells even though they do not produce a bona fide basement membrane-like layer.

A new procedure involving inhibition of a solid-phase radioimmunoassay was developed for specific determination of the outer-membrane protein and the lipopolysaccharide (LPS) serotypes of meningococci. Antigen was allowed to bind to the wells of a polyvinyl microtiter plate and then reacted with a limiting amount of homologous antibody which had been preincubated with buffer or a standard concentration of inhibiting antigen. The amount of antibody bound per well was quantitated by incubation with excess 125I-labeled goat anti-rabbit immunoglobulin. Typing sera for detecting eight LPS antigens and 18 protein antigens were made in rabbits by use of both the group C and group B bactericidal serotyping strains. Reactions between unabsorbed sera and purified LPS were inhibited in the LPS typing system, whereas reactions between absorbed sera and outer-membrane complex were inhibited in the protein typing system. Outer-membrane complex was used as the inhibiting antigen in both cases. Approximately 97% of the 80 group B and C strains tested were LPS typable, and 80% were protein typable. Of 51 group A strains tested, however, only 22% were LPS typable and 14% were protein typable. Several nonreciprocal correlations between the occurrence of particular LPS and protein serotype antigens on the same strain were observed, but in general the protein and LPS serotype antigens appeared to occur independently.

Thirty to fifty percent of group B and group C Neisseria meningitidis carrier isolates are not serotypable with existing outer membrane protein typing sera. A typing system based on differences in the outer membrane protein profiles after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was therefore developed as an adjunct to existing serotyping methods. Although most N. meningitidis strains contain several outer membrane proteins visible by SDS-PAGE, there are only one to three predominant proteins. The SDS-PAGE profiles of these major proteins were used to establish 10 different PAGE types. Greater than 95% of all meningococcal isolates, regardless of serogroup, fit into 1 of the 10 PAGE types. The outer membrane protein profile of individual strains after SDS-PAGE was constant when outer membrane fractions were prepared from the same strain on several different days. A comparison of gel profiles of meningococcal isolates obtained from different sites of the same patient revealed no significant differences among both major and minor proteins for isolate sets thus far examined. Characterization of strains by PAGE type can be a valuable epidemiological tool in addition to serotyping and in the absence of specific serotype antisera.

Careful analysis of affinity-purified class II molecules (Ia Ag) from the murine MHC revealed the existence of a set of associated molecules that consistently co-purified with the Ia Ag. SDS-PAGE revealed that molecules of Mr of 41 to 43 kDa and 56 to 58 kDa were associated with the affinity-purified I-Ak Ag from the AKR B cell lymphoma AKTB-1b. Two-dimensional electrophoresis (IEF vs SDS-PAGE) allowed further characterization of four molecules in the 41- to 43-kDa range and two in the 56- to 58-kDa range. All co-purifying proteins had isoelectric points between 5.2 and 6.2. The specificity of the association of the co-purifying molecules with the I-Ak Ag was established by using two criteria. First, with the exception of actin, proteins co-purifying with the I-Ak molecule were not found in samples of affinity-purified class I (H-2Kk) Ag or membrane Ig from the AKTB-1b lymphoma. Second, the use of the amino group-reactive homobifunctional cross-linker 3,3'-dithiobisproprionimidate with crude membranes from AKTB-1b increased the relative amount of materials co-purifying with I-Ak. The use of the membrane-impermeant cross-linker 3,3'-dithiobis(sulfosuccinimidyl) proprionate provided evidence that the interaction between I-Ak and one or more of the co-purifying components occurs on the cytoplasmic face of the membrane. Two of the co-purifying molecules have been identified. The major material in the 41- to 43-kDa range was partially sequenced, leading to its identification as cytoplasmic actin. One of the components in the 56- to 58-kDa range was tentatively identified as one of the isozymes (RII) of the regulatory subunit of the cAMP-dependent protein kinase, based on the use of the photoaffinity label 8-azido-cAMP.

Decay-accelerating factor (DAF) is a 70,000 Mr membrane protein that inhibits the amplification of the complement cascade on cell surfaces. Monoclonal antibodies against different epitopes of the 70,000 Mr DAF (DAF-1) recognize a second band at the position of 140,000 Mr on a Western blot of total red cell ghost proteins or partially pure DAF subjected to electrophoresis under denaturing conditions. Like DAF-1, this polypeptide (DAF-2) has the ability to accelerate decay of the C3 convertase, C4b2a, and to reincorporate into red cell membranes. A population of erythrocytes from patients with paroxysmal nocturnal hemoglobinuria (PNH) lack DAF-1 and also DAF-2. In addition, in some patients' red cells bearing DAF-1 of normal Mr, DAF-2 is 5,000 to 10,000 Mr smaller than normal. The structural basis for these differences in size of DAF and its PNH variants is unknown.

Whole cells and isolated outer membranes (OMs) of four strains of gonococci were surface radioiodinated with either lactoperoxidase or Iodogen (Pierce Chemical Co., Rockford, Ill.). These preparations were solubilized in sodium dodecyl sulfate and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Surface-radioiodinated protein I (PI) and PIII bands were excised from the sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and digested with alpha-chymotrypsin, and the resultant 125I-peptide fragments were resolved by high-voltage electrophoresis and thin-layer chromatography (i.e., surface peptide mapping). Radioemitting peptidic fragments were visualized by autoradiography. Results demonstrated that the PI molecule of each gonococcal strain studied had unique iodinatable peptides exposed on the surface of whole cells and OMs, whereas PIIIs appeared to have the same portion of the molecule exposed on the surface of bacteria or OMs, regardless of the gonococcal strain from which they were isolated. Many more radiolabeled peptides were seen in surface peptide maps of PIs from radiolabeled OMs than in those from radioiodinated whole cells, whereas different peptidic fragments were seen in the surface peptide maps of PIIIs from radiolabeled OMs than were seen in those from radiolabeled whole cells. These data suggest that PI may contribute strain-specific antigenic determinants and PIII may contribute cross-reactive determinants and that the surface exposure of PI and PIII is different in isolated OMs than in the OM of intact gonococci.

Polypeptide receptors on mammalian plasma membranes can be categorized on the basis of function. The binding of ligand by class I receptors results in changes in cell metabolism or behavior. Hormone-receptor interactions typify this group. The binding of ligand by class II receptors in ligand internalization. Although changes in cellular activity may result from metabolism of the internalized ligand, the interaction between ligand and class II receptor does not itself lead to alterations in cell behavior. Class II receptors include those for low-density lipoproteins and for alpha-macroglobulin-protease complexes. Although receptors within each category are chemically disparate, they show striking similarities in behavior. Analysis of the behavioral patterns of receptors in each category reveals insights into receptor physiology and allows for a prospective analysis of receptor characteristics.

The monoclonal antibody RB-8 heavily labels axons from the ventrolateral olfactory epithelium and their terminals in the glomeruli of the ventrolateral olfactory bulb, but leaves the axons from the dorsomedial epithelium unstained or lightly stained. RB-8 reacts with a 125 kDa membrane protein in both olfactory nerve and other parts of the CNS (Schwob and Gottlieb, 1986). Here we report further characterization of the molecular nature and cellular localization of the RB-8 antigen. The RB-8 antigen is exposed on the surface of olfactory axons. Individual axons and axon bundles stain when explant cultures of the fetal olfactory epithelium are incubated with monoclonal RB-8 antibody while living. The cell membrane is demonstrably intact, and access to the cell interior is blocked under these conditions, since the living axons do not stain if exposed to an antibody against a known intracellular constituent. The RB-8 antigen is an integral membrane protein. When assayed by direct radioimmunoassay (RIA), the antigen remains associated with brain membranes after extraction at pH 11, which solubilizes numerous other protein bands. The 125 kDa RB-8 antigen was purified to homogeneity from whole rat brains by extracting membranes with sodium deoxycholate, immunoaffinity chromatography over an RB-8 antibody column, and preparative one-dimensional SDS-PAGE. The NH2-terminal amino acid sequence is apparently unique among neuron-specific proteins that have been sequenced and has only an insignificant degree of homology with other known proteins. Two polyclonal rabbit antisera raised against the purified antigen recognize only the 125 kDa protein on immunoblots. Immunohistochemical staining of the primary olfactory projection with the antisera exactly matches that seen with monoclonal RB-8 antibody. Thus, the RB-8 antigens in brain and in olfactory nerve are highly homologous, if not identical. Furthermore, the results with the antisera suggest that the expression of the entire 125 kDa protein is regulated differentially between ventral and dorsal zones of the olfactory epithelium. The additional characterization of the RB-8 antigen reported here places constraints on the potential functions of this protein. The availability of polyclonal antisera may prove useful in assessing the role of this spatially segregated antigen in the primary olfactory projection.

Properties of human erythrocyte membrane proteins are widely studied recently. Comprehensive data were obtained on two main membrane proteins responsible for essential membrane functions. Besides, high performance two-dimensional electrophoresis and a number of other procedures enabled to detect more than 100 minor proteins in the biomembranes. The latest advances in studies of proteins and their coding genes allowed to compile a catalogue for erythrocyte membrane proteins, which is of importance for investigations in the field of membrane molecular biology. Structural organization of the catalogue for human erythrocyte membrane proteins is discussed.