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Biomedical subjects

H Meade

Publications and source records attributed to H Meade.

8 recordsLinked to original sources
PubMed1998

Transgenically produced human antithrombin: structural and functional comparison to human plasma-derived antithrombin.

Recombinant human antithrombin (rhAT) produced in transgenic goat milk was purified to greater than 99%. The specific activity of the rhAT was identical to human plasma-derived AT (phAT) in an in vitro thrombin inhibition assay. However, rhAT had a fourfold higher affinity for heparin than phAT. The rhAT was analyzed and compared with phAT by reverse phase high-performance liquid chromatography, circular dichroism, fluorophore-assisted carbohydrate electrophoresis (FACE), amino acid sequence, and liquid chromatography/mass spectrography peptide mapping. Based on these analyses, rhAT was determined to be structurally identical to phAT except for differences in glycosylation. Oligomannose structures were found on the Asn 155 site of the transgenic protein, whereas only complex structures were observed on the plasma protein. RhAT contained a GalNAc for galactose substitution on some N-linked oligosaccharides, as well as a high degree of fucosylation. RhAT was less sialylated than phAT and contained both N-acetylneuraminic and N-glycolylneuraminic acid. We postulate that the increase in affinity for heparin found with rhAT resulted from the presence of oligomannose-type structures on the Asn 155 glycosylation site and differences in sialylation.

Animals↗
PubMed1996

Alterations of the physical characteristics of milk from transgenic mice producing bovine kappa-casein.

kappa-Casein is the protein fraction of milk that allows formation of micelles and determines micelle size and function, thus affecting many of the physical characteristics of milk. Several lines of transgenic mice were generated bearing the B allele of the bovine kappa-CN gene under the control of the regulatory sequences of the caprine beta-CN gene that specifically directed expression of bovine kappa-CN to the lactating mammary tissue of these mice. High expression of bovine kappa-CN protein was observed in the lines studied; the total level of protein in milk was not significantly affected. A high degree of conservation in the amino acids involved in the predicted three-dimensional structure exists between murine and bovine kappa-CN. Milk from transgenic lines expressing high bovine kappa-CN had a significantly smaller micelle size than did control milk. Therefore, bovine kappa-CN appears to have effectively participated in assembly of murine casein micelles. There was no effect on the time of rennet coagulation, but the association was significant between the milk of transgenic lines and the production of a stronger curd in rennet-induced gels. We conclude that bovine kappa-CN is an appropriate candidate for transgenic technology that would increase the ratio of kappa-CN to the calcium-sensitive caseins, therefore affecting the physical properties of the colloidal casein suspension.

Amino Acid Sequence↗
PubMed1990

Bovine alpha S1-casein gene sequences direct high level expression of active human urokinase in mouse milk.

We have produced a line of transgenic mice carrying a hybrid bovine alpha S1 casein/human urokinase gene. Bovine alpha S1-casein gene regulatory sequences specifically direct expression of the human urokinase gene in lactating mammary tissue from these mice. Urokinase is a 54 kD protein with 9 disulfide bonds that is normally synthesized in the kidney; however, the casein/urokinase transgenic mice secrete active human urokinase into their milk at concentrations of 1-2 mg/ml. The mice show no other abnormalities. They give birth to, and nurse, normal sized healthy litters. Thus it is possible to produce high concentrations of a large, cysteine rich, non-milk protein in the milk of transgenic animals. This line of transgenic mice provides a model for the eventual production of transgenic farm animals producing high levels of recombinant proteins in their milk.

Animals↗