Document Detail


Expression of glutamate decarboxylase isoform, GAD65, in human mononuclear leucocytes: a possible implication of C-terminal end deletion by Western blot and RT-PCR study.
MedLine Citation:
PMID:  17965068     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Human peripheral blood leucocyte was examined for the expression of glutamate decarboxylase (GAD). Peripheral blood from healthy individuals was fractionated into polynuclear leucocytes and mononuclear leucocytes. Cell lysate from the mononuclear leucocytes was analysed by SDS-PAGE and Western blot. With antibody raised against unique C-terminal sequence for GAD65, two protein bands at 30 and 80 kDa were detected. However, with anti-GAD65/67 antibody recognizing very end of C-terminal, about 40 residues toward C-terminal, no protein bands were observed. Expression of mRNA coding for the epitope of these two antibodies was examined by using PCR technique. Results showed an evidence that mononuclear leucocytes express GAD65 with its C-terminal segment truncated. Our results have suggested an expression of GAD with the novel molecular weight may be caused by possible mononuclear leucocyte specific splicing errors.
Authors:
Satoko Matsukawa; Hiroshi Ueno
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Publication Detail:
Type:  Journal Article     Date:  2007-10-26
Journal Detail:
Title:  Journal of biochemistry     Volume:  142     ISSN:  0021-924X     ISO Abbreviation:  J. Biochem.     Publication Date:  2007 Nov 
Date Detail:
Created Date:  2007-11-16     Completed Date:  2008-04-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0376600     Medline TA:  J Biochem     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  633-8     Citation Subset:  IM    
Affiliation:
Laboratory of Applied Microbiology & Biochemistry, Nara Women's University, Nara 630-8506, Japan.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Antibodies / immunology
Blotting, Western / methods
Epitopes / immunology
Gene Expression Regulation, Enzymologic / genetics*
Glutamate Decarboxylase / blood,  chemistry,  genetics*
Humans
Isoenzymes / blood,  chemistry,  genetics,  metabolism*
Leukocytes, Mononuclear / metabolism*,  pathology
Molecular Sequence Data
Molecular Weight
Protein Isoforms / blood,  chemistry,  genetics,  metabolism*
RNA, Messenger / genetics,  metabolism
Reverse Transcriptase Polymerase Chain Reaction / methods
Chemical
Reg. No./Substance:
0/Antibodies; 0/Epitopes; 0/Isoenzymes; 0/Protein Isoforms; 0/RNA, Messenger; EC 4.1.1.15/Glutamate Decarboxylase

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