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Expression of a functional proteinase inhibitor capable of accepting xylose: bikunin

Falkenberg, Cecilia LU ; Wester Rosenlöf, Lena LU ; Belting, Mattias LU ; Eklund, Erik LU and Åkerström, Bo LU (2001) In Archives of Biochemistry and Biophysics 387(1). p.99-106
Abstract
Bikunin is a Kunitz-type proteinase inhibitor, which is cross-linked to heavy chains via a chondroitin sulfate chain, forming inter-alpha-inhibitor and related molecules. Rat bikunin was produced by baculovirus-infected insect cells. The protein could be purified with a total yield of 20 mg/liter medium. Unlike naturally occuring bikunin the recombinant protein had no galactosaminoglycan chain. Endoglycosidase digestion also suggested that the recombinant form lacked N-linked oligosaccharides. Bikunin is translated as a part of a precursor, alpha1-microglobulin/bikunin, but the functional significance of the cotranslation is unknown. Our results indicate that the proteinase inhibitory function of bikunin is not regulated by the... (More)
Bikunin is a Kunitz-type proteinase inhibitor, which is cross-linked to heavy chains via a chondroitin sulfate chain, forming inter-alpha-inhibitor and related molecules. Rat bikunin was produced by baculovirus-infected insect cells. The protein could be purified with a total yield of 20 mg/liter medium. Unlike naturally occuring bikunin the recombinant protein had no galactosaminoglycan chain. Endoglycosidase digestion also suggested that the recombinant form lacked N-linked oligosaccharides. Bikunin is translated as a part of a precursor, alpha1-microglobulin/bikunin, but the functional significance of the cotranslation is unknown. Our results indicate that the proteinase inhibitory function of bikunin is not regulated by the alpha1-microglobulin-part of the alpha1-microglobulin/bikunin precursor since recombinant bikunin had the same trypsin inhibitory activity as the recombinant precursor. Both free bikunin and the precursor were also functional as a substrate in an in vitro xylosylation system. This demonstrates that the alpha1-microglobulin-part is not necessary for the first step of galactosaminoglycan assembly. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
bikunin, greek small letter alpha1-microglobulin/bikunin precursor, proteinase inhibition, xylosylation, insect cells
in
Archives of Biochemistry and Biophysics
volume
387
issue
1
pages
99 - 106
publisher
Academic Press
external identifiers
  • pmid:11368189
  • scopus:0035263886
ISSN
0003-9861
DOI
10.1006/abbi.2000.2213
language
English
LU publication?
yes
id
531c46e9-e55e-4411-9f92-81efb580bcae (old id 1121578)
date added to LUP
2008-06-27 10:05:13
date last changed
2018-05-29 12:26:44
@article{531c46e9-e55e-4411-9f92-81efb580bcae,
  abstract     = {Bikunin is a Kunitz-type proteinase inhibitor, which is cross-linked to heavy chains via a chondroitin sulfate chain, forming inter-alpha-inhibitor and related molecules. Rat bikunin was produced by baculovirus-infected insect cells. The protein could be purified with a total yield of 20 mg/liter medium. Unlike naturally occuring bikunin the recombinant protein had no galactosaminoglycan chain. Endoglycosidase digestion also suggested that the recombinant form lacked N-linked oligosaccharides. Bikunin is translated as a part of a precursor, alpha1-microglobulin/bikunin, but the functional significance of the cotranslation is unknown. Our results indicate that the proteinase inhibitory function of bikunin is not regulated by the alpha1-microglobulin-part of the alpha1-microglobulin/bikunin precursor since recombinant bikunin had the same trypsin inhibitory activity as the recombinant precursor. Both free bikunin and the precursor were also functional as a substrate in an in vitro xylosylation system. This demonstrates that the alpha1-microglobulin-part is not necessary for the first step of galactosaminoglycan assembly.},
  author       = {Falkenberg, Cecilia and Wester Rosenlöf, Lena and Belting, Mattias and Eklund, Erik and Åkerström, Bo},
  issn         = {0003-9861},
  keyword      = {bikunin,greek small letter alpha1-microglobulin/bikunin precursor,proteinase inhibition,xylosylation,insect cells},
  language     = {eng},
  number       = {1},
  pages        = {99--106},
  publisher    = {Academic Press},
  series       = {Archives of Biochemistry and Biophysics},
  title        = {Expression of a functional proteinase inhibitor capable of accepting xylose: bikunin},
  url          = {http://dx.doi.org/10.1006/abbi.2000.2213},
  volume       = {387},
  year         = {2001},
}