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Localization of phosphatidylserine binding sites to structural domains of factor X-a

Srivastava, A; Wang, JF; Majumder, R; Rezaie, AR; Stenflo, Johan LU ; Esmon, CT and Lentz, BR (2002) In Journal of Biological Chemistry 277(3). p.1855-1863
Abstract
Binding of short chain phosphatidylserine (C6PS) enhances the proteolytic activity of factor X-a by 60-fold (Koppaka, V., Wang, J., Banerjee, M., and Lentz, B. R. (1996) Biochemistry 35, 7482-7491). In the present study, we locate three C6PS binding sites to different domains of factor X-a using a combination of activity, circular dichroism, fluorescence, and equilibrium dialysis measurements on proteolytic and biosynthetic fragments of factor X-a. Our results demonstrate that the structural responses of human and bovine factor X. to C6PS binding are somewhat different. Despite this difference, data obtained with fragments from both human and bovine factor X-a are consistent with a common hypothesis for the location of C6PS binding sites... (More)
Binding of short chain phosphatidylserine (C6PS) enhances the proteolytic activity of factor X-a by 60-fold (Koppaka, V., Wang, J., Banerjee, M., and Lentz, B. R. (1996) Biochemistry 35, 7482-7491). In the present study, we locate three C6PS binding sites to different domains of factor X-a using a combination of activity, circular dichroism, fluorescence, and equilibrium dialysis measurements on proteolytic and biosynthetic fragments of factor X-a. Our results demonstrate that the structural responses of human and bovine factor X. to C6PS binding are somewhat different. Despite this difference, data obtained with fragments from both human and bovine factor X-a are consistent with a common hypothesis for the location of C6PS binding sites to different structural domains. First, the gamma-carboxyglutamic acid (Gla) domain binds C6PS only in the absence of Ca2+ (k(d) similar to 1 mm), although this PS site does not influence the functional response of factor X-a. Second, a Ca2+-dependent binding site is in the epidermal growth factor domains (EGF(NC)) that are linked by Ca2+ and C6PS binding to the Gla domain. This site appears to be the lipid regulatory site of factor X-a. Third, a Ca2+-requiring site seems to be in the EGF(C)-catalytic domain. This site appears not to be a lipid regulatory site but rather to share residues with the substrate recognition site. Finally, the full functional response to C6PS requires linkage of the Gla, EGF(NC), and catalytic domains in the presence of Ca2+, meaning that PS regulation of factor X-a involves linkage between widely separated parts of the protein. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
277
issue
3
pages
1855 - 1863
publisher
ASBMB
external identifiers
  • wos:000173421300032
  • pmid:11707438
  • scopus:0037127212
ISSN
1083-351X
DOI
10.1074/jbc.M105697200
language
English
LU publication?
yes
id
d140eeb6-3307-4b82-bc7b-f818d04ff64d (old id 344437)
date added to LUP
2007-11-09 12:09:31
date last changed
2017-01-01 05:15:11
@article{d140eeb6-3307-4b82-bc7b-f818d04ff64d,
  abstract     = {Binding of short chain phosphatidylserine (C6PS) enhances the proteolytic activity of factor X-a by 60-fold (Koppaka, V., Wang, J., Banerjee, M., and Lentz, B. R. (1996) Biochemistry 35, 7482-7491). In the present study, we locate three C6PS binding sites to different domains of factor X-a using a combination of activity, circular dichroism, fluorescence, and equilibrium dialysis measurements on proteolytic and biosynthetic fragments of factor X-a. Our results demonstrate that the structural responses of human and bovine factor X. to C6PS binding are somewhat different. Despite this difference, data obtained with fragments from both human and bovine factor X-a are consistent with a common hypothesis for the location of C6PS binding sites to different structural domains. First, the gamma-carboxyglutamic acid (Gla) domain binds C6PS only in the absence of Ca2+ (k(d) similar to 1 mm), although this PS site does not influence the functional response of factor X-a. Second, a Ca2+-dependent binding site is in the epidermal growth factor domains (EGF(NC)) that are linked by Ca2+ and C6PS binding to the Gla domain. This site appears to be the lipid regulatory site of factor X-a. Third, a Ca2+-requiring site seems to be in the EGF(C)-catalytic domain. This site appears not to be a lipid regulatory site but rather to share residues with the substrate recognition site. Finally, the full functional response to C6PS requires linkage of the Gla, EGF(NC), and catalytic domains in the presence of Ca2+, meaning that PS regulation of factor X-a involves linkage between widely separated parts of the protein.},
  author       = {Srivastava, A and Wang, JF and Majumder, R and Rezaie, AR and Stenflo, Johan and Esmon, CT and Lentz, BR},
  issn         = {1083-351X},
  language     = {eng},
  number       = {3},
  pages        = {1855--1863},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Localization of phosphatidylserine binding sites to structural domains of factor X-a},
  url          = {http://dx.doi.org/10.1074/jbc.M105697200},
  volume       = {277},
  year         = {2002},
}