豌豆种子中3-磷酸甘油醛脱氢酶（GAPDH）的纯化与特性分析
Purification and characterization of glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) from pea seeds.
作者信息
Gani Zahid, Boradia Vishant Mahendra, Raghu Ram Janaki, Suryavanshi Prashant Mohan, Patil Pravinkumar, Kumar Santosh, Singh Ranvir, Raje Manoj, Raje Chaaya Iyengar
机构信息
National Institute of Pharmaceutical Education and Research, Phase X, Sector 67, SAS Nagar, Punjab, 160067, India.
Institute of Microbial Technology, Sector 39 A, Chandigarh, 160037, India.
出版信息
Protein Expr Purif. 2016 Nov;127:22-27. doi: 10.1016/j.pep.2016.06.014. Epub 2016 Jul 4.
Glyceraldehyde-3-phosphate dehydrogenase [GAPDH, NAD + oxidoreductase (phosphorylating) 1.2.1.12] catalyzes the conversion of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate coupled with the reduction of NAD(+) to NADH. In addition to its role in glycolysis, this enzyme has numerous alternate functions, in both prokaryotes and eukaryotes. In plants, additional functions have been reported from multiple species including Pisum sativum. A recent study has identified that GAPDH may play an important role in seed ageing and programmed cell death. Despite this the existing purification protocols are almost 40 years old, and only partial characterization of the enzyme has been reported. In the current study, we report a modified method for purification of enzymatically active pea seed GAPDH along with the characterization of the enzyme. Using 2D gel electrophoresis our study also demonstrates that pea seeds contain four isoforms of NAD(+) dependent GAPDH.
甘油醛-3-磷酸脱氢酶[GAPDH，NAD⁺氧化还原酶（磷酸化）1.2.1.12]催化甘油醛-3-磷酸转化为1,3-二磷酸甘油酸，并伴随着NAD⁺还原为NADH。除了在糖酵解中的作用外，这种酶在原核生物和真核生物中都有许多其他功能。在植物中，包括豌豆在内的多个物种都报道了其额外的功能。最近的一项研究发现，GAPDH可能在种子老化和程序性细胞死亡中起重要作用。尽管如此，现有的纯化方案已有近40年历史，且仅报道了该酶的部分特性。在本研究中，我们报告了一种纯化具有酶活性的豌豆种子GAPDH的改良方法以及该酶的特性。通过二维凝胶电泳，我们的研究还表明豌豆种子含有四种NAD⁺依赖性GAPDH同工型。
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