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Zinc-dependent conformational changes in domain D5 of high molecular mass kininogen modulate contact activation

Herwald, Heiko LU ; Mörgelin, Matthias LU ; Svensson, Henrik G. LU and Sjöbring, Ulf LU (2001) In European Journal of Biochemistry 268(2). p.396-404
Abstract

Human high molecular mass kininogen (HK) participates as nonenzymatic cofactor in the contact system. Here, we show that recombinant domain D5 of HK (rD5) prolongs the clotting time of the intrinsic pathway of coagulation and attenuates the generation of bradykinin. Further studies indicate that a correct fold of domain D5 within HK is required for the activation of the contact system. The folding of rD5 seems to be modulated by the metal ions Zn2+, Ni2+, and Cu2+ as a specific antibody directed against the zinc-binding site in HK binds to HK and rD5 in a metal ion concentration dependent manner. The finding that these three metal ions specifically affect contact activation suggests that they regulate... (More)

Human high molecular mass kininogen (HK) participates as nonenzymatic cofactor in the contact system. Here, we show that recombinant domain D5 of HK (rD5) prolongs the clotting time of the intrinsic pathway of coagulation and attenuates the generation of bradykinin. Further studies indicate that a correct fold of domain D5 within HK is required for the activation of the contact system. The folding of rD5 seems to be modulated by the metal ions Zn2+, Ni2+, and Cu2+ as a specific antibody directed against the zinc-binding site in HK binds to HK and rD5 in a metal ion concentration dependent manner. The finding that these three metal ions specifically affect contact activation suggests that they regulate the accessibility of rD5 for negatively charged surfaces. Support for the assumption that the observed phenomena are due to conformational changes was obtained by fluorescence spectroscopy of rD5, demonstrating that its fluorescence spectrum was changed in the presence of ZnCl2. Moreover, negative staining electron microscopy experiments suggest that the zinc-induced changes in D5 also affect the conformation of the entire HK protein. The present data emphasize the role of zinc and other metal ions in the regulation of contact activation.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Activated partial thromboplastin time, Bradykinin, Contact system, HK, Zinc
in
European Journal of Biochemistry
volume
268
issue
2
pages
396 - 404
publisher
Wiley-Blackwell
external identifiers
  • pmid:11168375
  • scopus:0034825602
ISSN
0014-2956
DOI
10.1046/j.1432-1033.2001.01888.x
language
English
LU publication?
yes
id
a26f6fc2-a49e-45b6-9976-428be30c4165
date added to LUP
2019-12-10 20:26:56
date last changed
2021-03-03 04:37:32
@article{a26f6fc2-a49e-45b6-9976-428be30c4165,
  abstract     = {<p>Human high molecular mass kininogen (HK) participates as nonenzymatic cofactor in the contact system. Here, we show that recombinant domain D5 of HK (rD5) prolongs the clotting time of the intrinsic pathway of coagulation and attenuates the generation of bradykinin. Further studies indicate that a correct fold of domain D5 within HK is required for the activation of the contact system. The folding of rD5 seems to be modulated by the metal ions Zn<sup>2+</sup>, Ni<sup>2+</sup>, and Cu<sup>2+</sup> as a specific antibody directed against the zinc-binding site in HK binds to HK and rD5 in a metal ion concentration dependent manner. The finding that these three metal ions specifically affect contact activation suggests that they regulate the accessibility of rD5 for negatively charged surfaces. Support for the assumption that the observed phenomena are due to conformational changes was obtained by fluorescence spectroscopy of rD5, demonstrating that its fluorescence spectrum was changed in the presence of ZnCl<sub>2</sub>. Moreover, negative staining electron microscopy experiments suggest that the zinc-induced changes in D5 also affect the conformation of the entire HK protein. The present data emphasize the role of zinc and other metal ions in the regulation of contact activation.</p>},
  author       = {Herwald, Heiko and Mörgelin, Matthias and Svensson, Henrik G. and Sjöbring, Ulf},
  issn         = {0014-2956},
  language     = {eng},
  number       = {2},
  pages        = {396--404},
  publisher    = {Wiley-Blackwell},
  series       = {European Journal of Biochemistry},
  title        = {Zinc-dependent conformational changes in domain D5 of high molecular mass kininogen modulate contact activation},
  url          = {http://dx.doi.org/10.1046/j.1432-1033.2001.01888.x},
  doi          = {10.1046/j.1432-1033.2001.01888.x},
  volume       = {268},
  year         = {2001},
}