The study on the structure of deoxyhypusine synthase in Saccharomyces cerevisiae
10.19405/j.cnki.issn1000-1492.2023.12.002
- VernacularTitle:酿酒酵母中脱氧羟腐胺赖氨酸合酶的结构研究
- Author:
Zhenzhen LI
1
;
Xiaoxiao MENG
;
Zhi QIAO
;
Li DAI
;
Xiaona YANG
;
Yanbin TENG
Author Information
1. 安徽医科大学生命科学学院,合肥 230032
- Keywords:
Saccharomyces cerevisiae;
deoxyhypusine synthase;
protein expression and purification;
crystal struc-ture;
X-ray diffraction
- From:
Acta Universitatis Medicinalis Anhui
2023;58(12):1995-2001
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the structure of deoxyhypusine synthase(DHS)in Saccharomyces cerevisiae(Dys1)and unravel the molecular mechanism of hypusine lysine modification,providing a theoretical basis for the treatment of highly proliferative diseases such as human immunodeficiency virus type 1(HIV-1)replication.Meth-ods Using the E.coli BL21 expression system,an in vitro expression vector was constructed and used to express the protein of Dys1.Dys1 protein samples were purified using methods such as affinity chromatography and molecu-lar sieving to achieve protein purification and isolation.The crystals of Dys1 were obtained using the crystallized so-lution containing 6％Polyethylene Glycol(PEG)8000,0.1 mol/L N-2-hydroxyethylpiperazine-N-ethane-sulphoni-cacid(Hepes)pH 6.5,and 8％ethylene glycol.The crystal structure of Dys1 was resolved at a resolution of 2.8 ? using X-ray crystallography.The structural analysis was performed with CCP4i and Coot software.Results The overall structure of Dys1 was a tetramer,each monomer containing a catalytic site and a cofactor NAD+binding site.The core region of the monomer adopted a Rossmann fold.The amino acid residues involved in the substrate binding sites were highly conserved among eukaryotes.Conclusion The crystal structure of Dys1 is being resolved for the first time.It reveals the binding mode of the cofactor NAD+to the enzyme and confirms that the enzyme functions as a tetramer,with the N-terminus serving as an essential modulator for its catalytic activity.
