Human tissue factor contains thioester-linked palmitate and stearate on the cytoplasmic half-cystine

Journal Article · Tue Jun 14 04:00:00 EDT 1988 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00412a004· OSTI ID:6904801

Bach, R; Konigsberg, W H; Nemerson, Y

The state of the five half-cystine residues in human tissue factor (TF) has been characterized. The results indicate that the four half-cystines in the extracellular domain of TF form two disulfide bonds and the half-cystine in the cytoplasmic region is acylated by plamitic acid and stearic acid. The extracellular disulfide cross-links, Cys/sup 49/-Cys/sup 57/ and Cys/sup 186/-Cys/sup 209/, were deduced from the analysis of tryptic peptides. Acylation of the cytoplasmic half-cystine was demonstrated by purifying and characterizing fibroblast TF from cells labeled with (/sup 3/H)palmitic acid. Radiolabeled fibroblast TF was observed by autoradiography following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The tritiated material covalently bound to the protein was identified as (/sup 3/H)palmitate and (/sup 3/H)stearate by reverse-phase high-pressure liquid chromatography. Deacylation of TF with hydroxylamine resulted in the spontaneous generation of disulfide-linked TF dimers. This result suggests that the disulfide-linked TF dimer, a minor component of most TF preparations, and the recently described heterodimeric form of TF are artifacts produced by deacylation of Cys/sup 245/ and subsequent interchain disulfide bond formation.

• Research Organization: Mt. Sinai School of Medicine of the City Univ. of New York, NY (USA)
• OSTI ID: 6904801
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 27:12; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
AMINO ACIDS
ANIMAL CELLS
ANIMALS
AUTORADIOGRAPHY
BODY
BRAIN
CARBON 14 COMPOUNDS
CARBOXYLIC ACID SALTS
CARBOXYLIC ACIDS
CENTRAL NERVOUS SYSTEM
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
CYSTINE
DISULFIDES
ELECTROPHORESIS
FIBROBLASTS
HEXADECANOIC ACID
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MAN
MOLECULAR STRUCTURE
MONOCARBOXYLIC ACIDS
NERVOUS SYSTEM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PRIMATES
PROTEINS
SEPARATION PROCESSES
SOMATIC CELLS
STEARATES
TRITIUM COMPOUNDS
VERTEBRATES