Mougin A, Corbier C, Soukri A, Wonacott A, Branlant C, Branlant G
Laboratoire d'Enzymologie et de Génie Génétique, UA CNRS 457, Faculté des Sciences, Vandoeuvre-les-Nancy, France.
Protein Eng. 1988 Apr;2(1):45-8. doi: 10.1093/protein/2.1.45.
Oligonucleotide-directed mutagenesis was employed to produce mutants of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of Escherichia coli and Bacillus stearothermophilus. Three different mutants proteins--His176----Asn, Cys149----Ser, Cys149----Gly--were isolated from one or both of the enzymes. The study of the properties of these mutants has shown that Cys149 is clearly responsible for the information of a charge-transfer transition, named the Racker band, observed during the NAD+ binding to apoGAPDH. This result excludes a similarity between the Racker band and the charge-transfer transition observed following the alkylation of GAPDH by 3-chloroacetyl pyridine-adenine dinucleotide.
采用寡核苷酸定向诱变技术制备大肠杆菌和嗜热脂肪芽孢杆菌甘油醛-3-磷酸脱氢酶（GAPDH）的突变体。从其中一种或两种酶中分离出三种不同的突变蛋白——His176→Asn、Cys149→Ser、Cys149→Gly。对这些突变体性质的研究表明，Cys149显然是NAD⁺与脱辅基GAPDH结合过程中观察到的一种名为Racker带的电荷转移跃迁信息的原因。该结果排除了Racker带与3-氯乙酰吡啶-腺嘌呤二核苷酸对GAPDH烷基化后观察到的电荷转移跃迁之间的相似性。
