Clostridium beijerinckii glyceraldehyde-3-phosphate dehydrogenase GAPDH: purification, crystallization, and X-ray crystallographic analysis

• 영문명: Clostridium beijerinckii glyceraldehyde-3-phosphate dehydrogenase GAPDH: purification, crystallization, and X-ray crystallographic analysis
• 발행기관: 한국구조생물학회
• 저자명: Jie Zhang Yuanyuan Chen Tingting Bu Xue Bai Shanru He Lulu Wang Chunshan Quan
• 간행물 정보: 『Biodesign』Vol 10, No 3, Sep, 51~55쪽, 전체 5쪽
• 주제분류: 자연과학 > 생물학
• 파일형태: PDF
• 발행일자: 2022.09.30

영문 초록

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a ubiquitous and highly abundant glycolytic enzyme. It plays a pivotal role for the energy and carbon metabolism of most organisms including industrial bacteria. It catalyzes the two step oxidative phosphorylation of D-glyceraldehyde-3-phosphate into 1,3-bisphosphoglycerate using inorganic phosphate and nicotinamide adenine dinucleotide (NAD+) as cofactor. In this study, GAPDH from C. beijerinckii (CbGAPDH) was successfully expressed and purified using Ni-NTA affinity, Q anion-exchange, and gel-filtration chromatography. The protein crystal was obtained and diffracted to a resolution of 1.60 Å. The crystal belonged to the hexagonal space group P6222, with unit-cell parameters of a = 120.6, b = 120.6, and c = 122.1 Å. The Matthews coefficient and solvent content were estimated to be 3.50 Å3 Da–1 and 64.90%, respectively, assuming that the asymmetric unit contained only one recombinant protein molecule.