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Biochemical characterization of glyceraldehyde-3-phosphate dehydrogenase from Thermococcus kodakarensis KOD1.
MedLine Citation:
PMID:  21409597     Owner:  NLM     Status:  Publisher    
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) plays an essential role in glycolysis by catalyzing the conversion of D: -glyceraldehyde 3-phosphate (D: -G3P) to 1,3-diphosphoglycerate using NAD(+) as a cofactor. In this report, the GAPDH gene from the hyperthermophilic archaeon Thermococcus kodakarensis KOD1 (GAPDH-tk) was cloned and the protein was purified to homogeneity. GAPDH-tk exists as a homotetramer with a native molecular mass of 145 kDa; the subunit molecular mass was 37 kDa. GAPDH-tk is a thermostable protein with a half-life of 5 h at 80-90°C. The apparent K (m) values for NAD(+) and D: -G3P were 77.8 ± 7.5 μM and 49.3 ± 3.0 μM, respectively, with V (max) values of 45.1 ± 0.8 U/mg and 59.6 ± 1.3 U/mg, respectively. Transmission electron microscopy (TEM) and image processing confirmed that GAPDH-tk has a tetrameric structure. Interestingly, GAPDH-tk migrates as high molecular mass forms (~232 kDa and ~669 kDa) in response to oxidative stress.
Baolei Jia; Le Thuy Linh; Sangmin Lee; Bang Phuong Pham; Jinliang Liu; Hongyu Pan; Shihong Zhang; Gang-Won Cheong
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-3-16
Journal Detail:
Title:  Extremophiles : life under extreme conditions     Volume:  -     ISSN:  1433-4909     ISO Abbreviation:  -     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-3-16     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9706854     Medline TA:  Extremophiles     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
College of Plant Sciences, Jilin University, Changchun, 130-062, China.
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