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We have previously reported the presence of a 70 kDa insulin-like growth factor (IGF)-II-specific binding protein in chicken serum using Western ligand blotting approaches. In order to ascertain the identity of this 70 kDa IGF-II binding species, the protein has been purified from chicken serum using a combination of ion-exchange and gel-permeation chromatography. Interestingly, amino acid sequencing of the purified protein revealed that it has the same N-terminal sequence as chicken vitronectin (VN). The protein has the ability to specifically bind IGF-II and not IGF-I as determined by ligand blotting, cross-linking and competitive binding assay approaches. In addition, the protein binds 125I-des(l-6)-IGF-II, suggesting that the interaction with IGF-II is different to those with other characterized IGF-binding proteins. Importantly, we have ascertained that both human and bovine VN also specifically bind IGF-II. These results are particularly relevant in the light of the recent report that the urokinase-type plasminogen activator receptor, a protein that also binds VN, has been shown to associate with the cation-independent mannose-6-phosphate/GF-II receptor and suggest a possible role for IGF-II in cell adhesion and invasion.

By numerically modeling pelagic sedimentation as a diffusive process, we study sedimentation effects in terms of stochastic parameters including seafloor RMS height, abyssal hill spacing, and slope distribution. Understanding gained allows the construction of inverse problems to obtain information about sediment distribution and basement morphology from multibeam bathymetric data. Using maximum likelihood estimation to compare slope distribution functions calculated from data to those from filtered model topographies, we estimate average sediment thickness L, basement RMS height H, and a measure of sediment mobility kappa. We invert for these parameters for 3-29 Ma seafloor from the western flank of the Mid-Atlantic Ridge (MAR) near 25 degree S, 2-45 Ma seafloor from the western flank of the MAR near 26 deg S, and 2-40 Ma seafloor from the eastern flank of the MAR near 25 deg S, and 1-38 Ma seafloor from the western flank of the MAR near 35 deg S. Variations in L allow us to constrain sediment rain rate and the corrosivity of bottom waters to calcite since the Oligocene. We hypothesize that sediment rain rates during the early and middle Miocene averaged only 10-50 % of more recent rates. Variations in H suggest correlation between tectonic setting and topographic variability. A narrow range of kappa describes intrahill sedimentation patterns.

Pelagic sedimentation is the primary modifier of topography generated by ridge-associated volcanic and tectonic processes. This thesis represents an effort to understand the processes of, and the general distribution of, pelagic sedimentation on rough topography, particularly in the Atlantic Basin but with applications to the world ocean as a whole. This study utilizes a simple numerical model of sedimentation which, when applied to models of rough basement topography, allows us to study sedimentation effects in terms of commonly-measured stochastic parameters including seafloor RMS height, abyssal hill spacing, and slope distribution. We also address the effect of sediment compaction on seafloor morphology, and the impact of long-wavelength topography on stochastic measures of sedimented seafloor. Understanding gained allows the construction of inverse problems to obtain information about sediment distribution and basement morphology from multibeam bathymetric data in regimes where backscatter from rough, reflective basement highs obscures returns from wide-beam seismic systems. By using maximum likelihood estimation to compare slope distribution functions calculated from data to those from filtered model topographies, we estimate average sediment thickness L, basement RMS height H, and a measure of sediment mobility K. Using data from near-ridge surveys and off-axis transit lines, we invert for L, H, and K for 3-29 Ma seafloor from the western flank of the Mid-Atlantic Ridge (MAR) near 26°N, 2- 45 Ma seafloor from the western flank of the MAR near 26°S, 2-40 Ma seafloor from the eastern flank of the MAR near 25°S, and 1-38 Ma seafloor from the western flank of the MAR near 35°S. Variations in L with seafloor age allow us to constrain sediment rain rate and the corrosivity of bottom waters to calcite since the Oligocene. We hypothesize that sediment rain rates during much of the early and middle Miocene were only 10-50% of the average rate for the past -10 m.y. Var

A strain of Vibrio cholerae, which had been engineered to express high levels of the non-toxic B subunit (EtxB) of Escherichia coli heat-labile enterotoxin, was subjected to transposon (TnphoA) mutagenesis. Two chromosomal TnphoA insertion mutations of the strain were isolated that showed a severe defect in the amount of EtxB produced. The loci disrupted by TnphoA in the two mutant derivatives were cloned and sequenced, and this revealed that the transposon had inserted at different sites in the same gene. The open reading frame of the gene predicts a 200-amino-acid exported protein, with a Cys-X-X-Cys motif characteristic of thioredoxin, protein disulphide isomerase, and DsbA (a periplasmic protein required for disulphide bond formation in E. coli). The V. cholerae protein exhibited 40% identity with the DsbA protein of E. coli, including 90% identity in the region of the active-site motif. Introduction of a plasmid encoding E. coli DsbA into the V. cholerae TnphoA derivatives was found to restore enterotoxin formation, whilst expression of Etx or EtxB in a dsbA mutant of E. coli confirmed that DsbA is required for enterotoxin formation in E. coli. These results suggest that, since each EtxB subunit contains a single intramolecular disulphide bond, a transient intermolecular interaction with DsbA occurs during toxin subunit folding which catalyses formation of the disulphide in vivo.