Human Tissue Factor Contains Thioester-Linked Palmitate and Stearate on the Cytoplasmic Half-Cystine

Ronald R Bach, William H. Konigsberg, Yale Nemerson

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Abstract

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 palmitic acid and stearic acid. the extracellular disulfide cross-links, Cys49-Cys57and Cys186-Cys209, 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 [3H] 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 [3H] palmitate and [3H] stearate by reverse-phase high-pressure liquid chromatography. Deacylation of TF with hydroxylamine resulted in the spontaneous generation of di-sulfide-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 Cys245and subsequent interchain disulfide bond formation.

Original languageEnglish (US)
Pages (from-to)4227-4231
Number of pages5
JournalBiochemistry
Volume27
Issue number12
DOIs
StatePublished - Jun 1 1988

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