PMID- 22543501
OWN - NLM
STAT- MEDLINE
DCOM- 20120928
LR  - 20131121
IS  - 1745-7270 (Electronic)
IS  - 1672-9145 (Linking)
VI  - 44
IP  - 6
DP  - 2012 Jun
TI  - Purification and partial characterization of glyceraldehyde-3-phosphate 
      dehydrogenase from the ciliate Tetrahymena thermophila.
PG  - 527-34
LID - 10.1093/abbs/gms028 [doi]
AB  - In the present study, we purified the glycolytic enzyme 
      glyceraldehyde-3-phosphate dehydrogenase (GAPDH) which is involved in cellular 
      energy production and has important housekeeping functions, from the ciliate 
      Tetrahymena thermophila using a three-step procedure. The enzyme was purified ~68 
      folds by ammonium sulfate precipitation, followed by two steps of column 
      chromatography (DEAE-cellulose and Mono-S). The purified enzyme is a homotetramer 
      with a molecular weight of ~120 kDa. Isoelectric focusing analysis showed the 
      presence of only one basic GAPDH isoform with an isoelectric point of 8.8. 
      Western blot analysis showed a single 32-kDa band corresponding to the enzyme 
      subunit using a monospecific polyclonal antibody against the T. thermophila 
      GAPDH. The maximum of enzyme activity occurred at pH 8.0 and at 30-35 degrees C. The 
      apparent K(m) values for both NAD(+) and D-glyceraldehyde-3-phosphate were 0.102 
      +/- 0.012 and 0.360 +/- 0.018 mM, respectively. The maximal velocity (V(max)) was 
      39.40 +/- 2.95 U/mg. The T. thermophila GAPDH is inhibited by oxidative and 
      nitrosative stress reagents.
FAU - Errafiy, Nadia
AU  - Errafiy N
AD  - Laboratoire de Physiologie et Genetique Moleculaire, Departement de Biologie, 
      Faculte des Sciences Ain Chock, Universite Hassan II-Ain Chock, Km 8 Route d'El 
      Jadida, BP. 5366 Maarif, Casablanca, Morocco. naderlana@hotmail.com
FAU - Soukri, Abdelaziz
AU  - Soukri A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120427
PL  - China
TA  - Acta Biochim Biophys Sin (Shanghai)
JT  - Acta biochimica et biophysica Sinica
JID - 101206716
RN  - 169D1260KM (Nitroprusside)
RN  - BBX060AN9V (Hydrogen Peroxide)
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
SB  - IM
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/antagonists & inhibitors/*isolation & 
      purification/metabolism
MH  - Hydrogen Peroxide/pharmacology
MH  - Hydrogen-Ion Concentration
MH  - Isoelectric Focusing
MH  - Kinetics
MH  - Nitroprusside/pharmacology
MH  - Temperature
MH  - Tetrahymena thermophila/*enzymology
EDAT- 2012/05/01 06:00
MHDA- 2012/09/29 06:00
CRDT- 2012/05/01 06:00
PHST- 2012/05/01 06:00 [entrez]
PHST- 2012/05/01 06:00 [pubmed]
PHST- 2012/09/29 06:00 [medline]
AID - gms028 [pii]
AID - 10.1093/abbs/gms028 [doi]
PST - ppublish
SO  - Acta Biochim Biophys Sin (Shanghai). 2012 Jun;44(6):527-34. doi: 
      10.1093/abbs/gms028. Epub 2012 Apr 27.