Linked Life Data

9587405

Source:http://linkedlifedata.com/resource/pubmed/id/9587405

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1998-6-3
pubmed:abstractText
Human blood coagulation factor VII has unique carbohydrate moieties O-glycosidically linked to serine 52 and serine 60 residues in its first epidermal growth factor-like domain. To study the functional role of these glycosyl moieties in factor VII, we constructed, expressed, and purified site-specific recombinant mutants of human factor VII in which serine 52 and serine 60 were conservatively replaced with alanine residues. S52A factor VIIa (Ser-52-->Ala), S60A factor VIIa (Ser-60-->Ala), and S52,60A factor VIIa (Ser-52, Ser-60-->Ala) exhibited 56, 73, and 44%, respectively, of the clotting activity of wild-type factor VIIa using human brain thromboplastin as a source of tissue factor/phospholipids and 32, 43, and 14% of wild-type factor VIIa using a mixture of recombinant soluble tissue factor and mixed brain phospholipids. The tissue factor-dependent and -independent amidolytic activities of these mutants were essentially indistinguishable from that of wild-type factor VIIa. In addition, equilibrium dialysis experiments indicated that the profiles of 45Ca2+ binding to these mutants were identical with that of wild-type factor VII. In the presence of either Ca2+ or EGTA, the Kd values for the interaction of the three factor VIIa mutants to full-length tissue factor were 2- to 5-fold higher than that of wild-type factor VIIa, while the Kd values for the interaction of these mutants to soluble tissue factor were 4- to 15-fold higher than that of wild-type factor VIIa. Measurement of the association and dissociation rate constants for factor VIIa binding to relipidated tissue factor apoprotein revealed that the association rate constants of the three factor VII mutants were decreased in comparison with that of wild-type factor VIIa, while the dissociation rate constants of these three mutants were virtually identical to that of wild-type factor VIIa. These findings strongly suggest that glycosyl moieties attached to Ser-52 and Ser-60 in factor VII/VIIa provide unique structural elements that are important for the rapid association of factor VII/VIIa with its cellular receptor and cofactor.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0003-9861
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
352
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
182-92
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1998
pubmed:articleTitle
Functional consequences of mutations in Ser-52 and Ser-60 in human blood coagulation factor VII.
pubmed:affiliation
Department of Pathology, University of New Mexico School of Medicine, Albuquerque 87131, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.