Tissue factor, soluble domain

We have experimented with Trp analog labeling protocols for recombinant proteins produced in bacteria and yeast, and we have found variability in these labeled proteins both with respect to structural and functional integrity and degree of analog incorporation (1,2). We have evaluated levels of 5-OHTrp incorporation in several protein systems. In this paper, we evaluate attempts to replace the four Trps in the truncated, soluble domain of recombinant human Tissue Factor (sTF) with 5-OHTrp and 7-ATrp. This domain of native, membrane-bound tissue factor binds factor Wa, a serine protease circulating in... 

Konigsberg WH, Rusinova E, Waxman E Testing an FTIR-Consistent Model of the Soluble Domain of Human Tissue Factor. ACS Symposium Series, in press... 

The epidermal growth factor (EGF) domain is one example of a module that is present in several proteins (Figure 3-8). EGF is a small, soluble peptide hormone that binds to cells in the embryo and in skin and connective tissue in adults, causing them to divide. It is generated by proteolytic cleavage between repeated EGF domains in the EGF precursor protein, which is anchored in the cell membrane by a membrane-spanning domain. EGF modules are also present in other proteins and are liberated by proteolysis these proteins include tissue plasminogen activator (TPA), a protease that is used to dissolve blood clots In heart attack victims ... 

Figure 4 Schematic representation of the polymerization of fibrin and fibrinolysis. Fibrinogen molecules are soluble in the blood until thrombin cleaves the A fibrinopeptides, yielding desA fibrin monomer, which assemble in a half-staggered fashion to make two-stranded protofibrils. Factor Xllla rapidly crosslinks the adjacent y chains. The B fibrinopeptides are cleaved more slowly mosdy after polymerization begins. On cleavage of the B fibrinopeptides, the aC domains are released, bringing protofibrils together more efficiendy to produce thicker fibers. Factor Xllla crosslinks the a chains more slowly, to stabilize the clot more fully. Plasminogen is converted to plasmin by tissue plasminogen activator on the fibrin surface plasmin cleaves specific peptide bonds (arrows) to dissolve the clot.

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