Arguments against a close relationship between non‐phosphorylating and phosphorylating glyceraldehyde‐3‐phosphate dehydrogenases

FEBS Letters

Riva, M.¹  Branlant, G.³  Carles, C.¹  Trost, P.²  Scagliarini, S.²  Seta, F.Della³  Michels, S.³ 

[1] Laboratoire d'Ingénierie des Protéines, Centre d'Etudes Nucléaires de Saclay, 91191 Gifsur- Yvette Cédex, France;Università degli Studi ai Bologna, Dipartimento di Biologia/Botanica, Via Irnerio 42, 40126 Bologna, Italy;Laboratoire d'Enzymologie et de Génie Génétique, Université de Nancy I, URA CNRS 457, BP 239, 54506 Vandoeuvre-lès-Nancy Cedex, France

关键词: Stereospecificity;    NMR;    Glyceraldehyde-3-phosphate dehydrogenase;    Spinach;    Evolution;    B. stearothermophilus;    Bacillus stearothermophilus;    E. coli;    Escherichia coli;    GAPDH;    glyceraldehyde-3-phosphate dehydrogenase;    ALDH;    aldehyde dehydrogenase;    1;    3 dPG;    1;    3-diphosphoglycerate;    G3P glyceraldehyde-3-phosphate;    3PG;    3-phosphoglycerate;    PTH-amino acids;    phenylthiohydantoin amino acids;   

DOI  :  10.1016/0014-5793(94)80393-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Non-phosphorylating NADP-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (EC 1.2.1.9) from spinach leaves was purified to homogeneity using an improved purification procedure. Thus, a major contaminant with molecular mass and ion-exchange properties similar to non-phosphorylating GAPDH was eliminated. Using this pure non-phosphorylating GAPDH, cofactor stereospecificity was determined by ¹H NMR. Analysis of the NADPH formed from the hydride transfer from glyceraldehyde-3-phosphate to [4-²H]NADP showed that the enzyme belongs to the A-stereospecific dehydrogenase family. This stereospecificity is the same as that described for the aldehyde dehydrogenase (ALDH) superfamily and opposite to that of the phosphorylating GAPDH. Moreover, results from peptide sequencing analysis suggest a similarity in sequence between the non-phosphorylating GAPDH and ALDHs. Thus, the results taken all together strongly suggest that non-phosphorylating GAPDH belongs to the ALDH family and has no close relationship to the phosphorylating GAPDH class.

【 授权许可】

Unknown