Our findings demonstrate that a minimal mutation at the NAD⁺-binding site is sufficient to generate a competitive but dysfunctional GAPDH, and its ectopic expression inhibits the wild-type to disrupt glycolysis.

既往研究表明，细胞质gapdh比核gapdh具有更高的糖酵解活性，上调细胞质gapdh的表达可增强CD8+ T细胞的糖酵解代谢并促进其激活。然而，作为gapdh亚细胞定位基础的分子事件是难以捉摸的。

2018年12月1日 · Here, we report that a catalytically-deficient mutant-GAPDH competitively inhibits the wild-type, and disrupts glucose metabolism in cancer cells. Using site-directed mutagenesis, the human GAPDH clone was mutated at one of the NAD + -binding sites, (i.e.) arginine (R13) and isoleucine (I14) to glutamine (Q13) and phenylalanine (F14), respectively.

This diagram represents possible feedback-loop mechanisms of p53 modulation by GAPDH and the NAD +-dependant protein deacetylase, Sirt-1. Upon binding the GAPDH-NAD + complex, Sirt-1 undergoes a conformational change that enables it to bind and remove acetyl groups (−OCH 3) from C-terminal Lys residues of p53, inactivating it. Inactivated p53 ...

2015年11月26日 · Site-mutagenesis at positions S98 and T99 in the NAD(+) binding center reduced enzymatic activity of GAPDH due to decreased affinity to NAD(+) (Km = 741 ± 257 μmol/L in T99I vs 57 ± 11.1 µmol/L in wild type GAPDH. Molecular modeling experiments revealed the effect of mutations on NAD(+) binding with GAPDH.

2014年12月1日 · Protein RNA interactions represent a fundamental mechanism underlying post-transcriptional gene regulation. Recent studies identified GAPDH as an AU 3′-UTR binding protein mediated by its RNA binding domain within the GAPDH NAD + binding site (Fig. 1). The latter was defined by peptide mapping and by NAD + competition (Nagy and Rigby, 1995).

2019年3月11日 · Our findings provide an integrated explanation for the dependence of inflammatory macrophages on the NAD + salvage pathway. Macrophages reside in mammalian tissues and can be differentially...

E. coli cells have NAD +-dependent glycolytic GAPDH. One reasonable approach to increase NADPH formation in cells is to change the specificity of the GAPDH from NAD + to NADP +. In this study, we modified the cofactor specificity of E. coli GAPDH by amino acid substitutions at positions 34, 188 and 189.

2020年3月13日 · In the present work, using human and murine cells; immunoprecipitation, pulldown, and surface plasmon resonance assays; and immunofluorescence, small-angle X-ray scattering, and MS-based analyses, we report that GAPDH and NAMPT form a stable complex that is essential for nuclear translocation of NAMPT.

These results indicate an unexpected, important role phosphorylated amino acids within the NAD + binding center play in intranuclear functions of GAPDH, and suggest a single mechanism by which GAPDH contributes to formation and functioning of its multiple binding partners.