Post-translational modifications of recombinant human tissue factor.
Recombinant human tissue factor (rTF) purified from transfected mammalian cells is a glycoprotein that contains N-linked, but not O-linked oligosaccharides. Two of the three potential N-linked sites in the extracellular portion are fully glycosylated, while one site is approximately 90% utilized. These sites have complex-type oligosaccharides attached. The potential N-linked site in the cytoplasmic domain near the C-terminus is not glycosylated. Characterization of the tryptic map of rTF confirmed most of the proposed amino acid sequence. In addition, the disulfide bonds (between Cys-49 and Cys-57 and between Cys-186 and Cys-209) were demonstrated by FAB-MS analysis of cysteine-containing fragments obtained from the tryptic map.
Paborsky
L R
LR
Department of Cardiovascular Research, Genentech, Inc., South San Francisco, CA 94080.
Harris
R J
RJ
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D002999:Clone Cells; D004220:Disulfides / Q000737:chemistry; D005260:Female; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D010920:Placenta / Q000737:chemistry; D011247:Pregnancy; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D011994:Recombinant Proteins / Q000737:chemistry; D013925:Thromboplastin / Q000737:chemistry*; D014357:Trypsin; D014415:Tunicamycin / Q000494:pharmacology
D004220:Disulfides; D011994:Recombinant Proteins; D014415:Tunicamycin; D013925:Thromboplastin; D014357:Trypsin