Biophysical and structural characterization of the thermostable WD40 domain of a prokaryotic protein, Thermomonospora curvata PkwA

Shen, Chen1; Du, Ye1,2; Qiao, Fangfang1; Kong, Tian1; Yuan, Lirong1; Zhang, Delin1; Wu, Xianhui1; Li, Dongyang1,3; Wu, Yun-Dong1,4

2018-08-28

10.1038/s41598-018-31140-y

Scientific Reports   影响因子和分区

2045-2322

WD40 proteins belong to a big protein family with members identified in every eukaryotic proteome. However, WD40 proteins were only reported in a few prokaryotic proteomes. Using WDSP (http://wu.scbb.pkusz.edu.cn/wdsp/), a prediction tool, we identified thousands of prokaryotic WD40 proteins, among which few proteins have been biochemically characterized. As shown in our previous bioinformatics study, a large proportion of prokaryotic WD40 proteins have higher intramolecular sequence identity among repeats and more hydrogen networks, which may indicate better stability than eukaryotic WD40s. Here we report our biophysical and structural study on the WD40 domain of PkwA from Thermomonospora curvata (referred as tPkwA-C). We demonstrated that the stability of thermophilic tPkwA-C correlated to ionic strength and tPkwA-C exhibited fully reversible unfolding under different denaturing conditions. Therefore, the folding kinetics was also studied through stopped-flow circular dichroism spectra. The crystal structure of tPkwA-C was further resolved and shed light on the key factors that stabilize its beta-propeller structure. Like other WD40 proteins, DHSW tetrad has a significant impact on the stability of tPkwA-C. Considering its unique features, we proposed that tPkwA-C should be a great structural template for protein engineering to study key residues involved in protein-protein interaction of a WD40 protein.

SCI

英语

通讯

Health and Family Planning Commission Project of Shenzhen Municipality, China[201606003]

Science & Technology - Other Topics

Multidisciplinary Sciences

WOS:000442917500037

NATURE PUBLISHING GROUP

Web of Science

1.Peking Univ, Shenzhen Grad Sch, Lab Chem Genom, Lab Computat Chem & Drug Design, Shenzhen 518055, Peoples R China
2.Peoples Hosp Longhua, Med Res Ctr, Shenzhen 518109, Peoples R China
3.Southern Univ Sci & Technol, SUSTech Acad Adv Interdisciplinary Studies, Shenzhen 518055, Peoples R China
4.Peking Univ, Coll Chem, Beijing 100871, Peoples R China

Shen, Chen,Du, Ye,Qiao, Fangfang,et al. Biophysical and structural characterization of the thermostable WD40 domain of a prokaryotic protein, Thermomonospora curvata PkwA[J]. Scientific Reports,2018,8.

Shen, Chen.,Du, Ye.,Qiao, Fangfang.,Kong, Tian.,Yuan, Lirong.,...&Wu, Yun-Dong.(2018).Biophysical and structural characterization of the thermostable WD40 domain of a prokaryotic protein, Thermomonospora curvata PkwA.Scientific Reports,8.

Shen, Chen,et al."Biophysical and structural characterization of the thermostable WD40 domain of a prokaryotic protein, Thermomonospora curvata PkwA".Scientific Reports 8(2018).