Function-Biased Choice of Additives for Optimization of Protein Crystallization: The Case of the Putative Thioesterase PA5185 from Pseudomonas aeruginosa PAO1

详细信息    查看全文

• 作者：Maksymilian Chruszcz ; Matthew D. Zimmerman ; Shuren Wang ; Katarzyna D. Koclega ; Heping Zheng ; Elena Evdokimova ; Marina Kudritska ; Marcin Cymborowski ; Alexei Savchenko ; Aled Edwards ; Wladek Minor
• 刊名：Crystal Growth & Design
• 出版年：2008
• 出版时间：November 5, 2008
• 年：2008
• 卷：8
• 期：11
• 页码：4054-4061
• 全文大小：437K
• 年卷期：v.8,no.11(November 5, 2008)
• ISSN：1528-7505

文摘

The crystal structure of PA5185, a putative thioesterase from Pseudomonas aeruginosa strain PAO1, was solved using multi-wavelength anomalous diffraction to 2.4 Å. Analysis of the structure and information about the putative function of the protein were used to optimize crystallization conditions. The crystal growth was optimized by applying additives with chemical similarity to a fragment of a putative PA5185 substrate (CoA or its derivative). Using new crystallization conditions containing this function-biased set of additives, several new crystal forms were produced, and structures of three of them (in three different space groups) were determined. One of the new crystal forms had an improved resolution limit of 1.9 Å, and another displayed an alternative conformation of the highly conserved loop containing Asn26, which could play a physiological role. Surprisingly, none of the additives were ordered in the crystal structures. Application of function-biased additives could be used as a standard optimization protocol for producing improved diffraction, or new crystal forms, which may lead to better understanding of the biological functions of proteins.