The third Trp-Lys-Ser (WKS) tripeptide motif in tissue factor is associated with a function site

The tripeptide sequence Trp-Lys-Ser (WKS) is repeated three times in the extracellular ligand binding domain of human Tissue Factor (TF). Using site-directed mutagenesis, we replaced each of the WKS motifs in human TF by Arg-Lys-Gly (RKG), the least conserved replacement for the motif found in murin...

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Bibliographic Details
Published in:Biochemical journal 1992-03, Vol.282 (3), p.737-740
Main Authors: REHEMTULLA, A, RUF, W, MILES, D. J, EDGINGTON, T. S
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Binding Sites
Biological and medical sciences
Blood coagulation. Blood cells
CHO Cells
Coagulation factors
Cricetinae
Cricetulus
Exons - genetics
Extracellular Space - physiology
Fundamental and applied biological sciences. Psychology
Humans
Mice
Molecular and cellular biology
Molecular Sequence Data
Mutagenesis, Site-Directed
Protein Conformation
Rabbits
Repetitive Sequences, Nucleic Acid - genetics
Repetitive Sequences, Nucleic Acid - physiology
Sequence Homology, Nucleic Acid
Thromboplastin - chemistry
Thromboplastin - genetics
Thromboplastin - physiology
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Summary:The tripeptide sequence Trp-Lys-Ser (WKS) is repeated three times in the extracellular ligand binding domain of human Tissue Factor (TF). Using site-directed mutagenesis, we replaced each of the WKS motifs in human TF by Arg-Lys-Gly (RKG), the least conserved replacement for the motif found in murine TF. This substitution in the first repeat W14KS, as well as a Trp14---Arg substitution, resulted in a structurally altered protein, whereas a conservative hydrophobic Trp14---Phe substitution resulted in a functionally normal protein. This suggests that Trp14 may contribute to a hydrophobic core rather than involvement of this motif in function. Replacement of the W45KS and W158KS motifs was associated with no detectable structural alterations; however, function was diminished with the RKG replacement of the third repeat. Mutant proteins with Lys159---Ala and Tyr157---Ala substitutions exhibited loss of function, whereas Tyr156---Ala and Ser160---Ala substitutions flanking the YWK sequence resulted in functional proteins. These data demonstrate that the W158KS motif in human TF is associated with a functional site and identify Lys159 in this motif as a functionally important residue.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj2820737