Linked Life Data

3091068

Source:http://linkedlifedata.com/resource/pubmed/id/3091068

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
14
pubmed:dateCreated
1986-10-23
pubmed:abstractText
One way in which coagulation may be initiated is by the action of factor VIIa (a plasma serine protease) and tissue factor (a membrane-bound lipid-dependent glycoprotein). We show that in the absence of either factor VIIa or tissue factor, the activation of the natural coagulation substrates, factors IX and X, is not detectable; i.e., tissue factor is an essential activator. We propose that the reaction is fully ordered; that is, the enzyme-activator complex picks up substrate to form a ternary product forming species. Our model precludes the formation of enzyme-substrate and activator-substrate complexes. We have derived equations for the two possible variations of this model: one in which product formation is accompanied by the release of the enzyme-activator complex and the other in which product, free enzyme, and free activator are formed with each catalytic cycle. Our data support only the former which is consistent with both steady-state and rapid equilibrium assumptions. The model is supported by experiments using a monoclonal anti-tissue factor antibody, which affects only the Km app, and a modified form of factor VIIa, which, depending on the sequence in which reagents are added to the reaction, either decreases the Vmax or increases the Km app. We present equations describing the initial velocity of these reactions. Utilizing dilution-jump experiments, we show that the system is hysteretic and suggest that this phenomenon is due to a slow release of enzyme from activator. However, the kinetically determined dissociation constant of enzyme and activator, previously found to be 4.5 nM under equilibrium conditions, was estimated to be 0.04-0.09 nM. Accordingly, we examined other essential activation models in which the product-forming species consists of a complex of enzyme, activator, and substrate at a molar ratio of 1:1:1; none could account for the apparent tight binding of enzyme and activator. We therefore postulate an ordered addition, essential activation model in which the enzyme undergoes two conformational transformations: one as a consequence of binding to tissue factor, resulting in a species which binds to and hydrolyzes its natural substrates. The other conformational change in the enzyme is induced by substrate, resulting in a species which binds more tightly to its activator. Thus, we hypothesize a "conformational cage" which precludes the dissociation of enzyme from activator while significant concentrations of substrate are present.
pubmed:grant
pubmed:commentsCorrections
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
25
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4020-33
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1986
pubmed:articleTitle
An ordered addition, essential activation model of the tissue factor pathway of coagulation: evidence for a conformational cage.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't