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Site-directed mutagenesis of K396R of the 65 kDa glutamic acid decarboxylase active site obliterates enzyme activity but not antibody binding

Hampe, Christiane S; Hammerle, Lisa P; Falorni, Alberto; Robertson, John D. and Lernmark, Åke LU (2001) In FEBS Letters 488(3). p.185-189
Abstract

The role of K396 in the enzymatic catalysis and the antigenicity of the 65 kDa isoform of glutamate decarboxylase (GAD65) was analyzed using the K396R GAD65 mutant. GAD65 is a major autoantigen in Type 1 diabetes and autoantibodies directed to GAD65 are widely used markers for this disease. We found that (1) recombinant human GAD65 is fully enzymatically active; (2) the K396R mutation abolished GAD65 activity; and (3) the K396R mutant retained full antigenicity to GAD65 autoantibodies in serum from Type 1 diabetes patients, but not to polyclonal antibodies raised to the catalytic domain.

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author
publishing date
type
Contribution to journal
publication status
published
keywords
Autoimmunity, Diabetes, Glutamate decarboxylase, Pyridoxal 5-phosphate
in
FEBS Letters
volume
488
issue
3
pages
5 pages
publisher
Wiley-Blackwell
external identifiers
  • scopus:0035910379
ISSN
0014-5793
DOI
10.1016/S0014-5793(00)02429-7
language
English
LU publication?
no
id
ea02044e-5b96-429a-94fc-5c25438d148d
date added to LUP
2017-09-07 09:05:36
date last changed
2018-05-29 11:51:13
@article{ea02044e-5b96-429a-94fc-5c25438d148d,
  abstract     = {<p>The role of K396 in the enzymatic catalysis and the antigenicity of the 65 kDa isoform of glutamate decarboxylase (GAD65) was analyzed using the K396R GAD65 mutant. GAD65 is a major autoantigen in Type 1 diabetes and autoantibodies directed to GAD65 are widely used markers for this disease. We found that (1) recombinant human GAD65 is fully enzymatically active; (2) the K396R mutation abolished GAD65 activity; and (3) the K396R mutant retained full antigenicity to GAD65 autoantibodies in serum from Type 1 diabetes patients, but not to polyclonal antibodies raised to the catalytic domain.</p>},
  author       = {Hampe, Christiane S and Hammerle, Lisa P and Falorni, Alberto and Robertson, John D. and Lernmark, Åke},
  issn         = {0014-5793},
  keyword      = {Autoimmunity,Diabetes,Glutamate decarboxylase,Pyridoxal 5-phosphate},
  language     = {eng},
  month        = {01},
  number       = {3},
  pages        = {185--189},
  publisher    = {Wiley-Blackwell},
  series       = {FEBS Letters},
  title        = {Site-directed mutagenesis of K396R of the 65 kDa glutamic acid decarboxylase active site obliterates enzyme activity but not antibody binding},
  url          = {http://dx.doi.org/10.1016/S0014-5793(00)02429-7},
  volume       = {488},
  year         = {2001},
}