用于蛋白质核磁共振光谱的聚丙烯酰胺稳定的Pfl噬菌体液晶的表征

Characterization of polyacrylamide-stabilized Pfl phage liquid crystals for protein NMR spectroscopy.

作者信息

Trempe Jean-François, Morin Frederick G, Xia Zhicheng, Marchessault Robert H, Gehring Kalle

机构信息

Department of Biochemistry and bDepartment of Chemistry, McGill University, Montreal, QC, Canada.

出版信息

J Biomol NMR. 2002 Jan;22(1):83-7. doi: 10.1023/a:1013832422428.

DOI:10.1023/a:1013832422428

PMID:11885983

Abstract

A new polymer-stabilized nematic liquid crystal has been characterized for the measurement of biomolecular residual dipolar couplings. Filamentous Pf1 phage were embedded in a polyacrylamide matrix that fixes the orientation of the particles. The alignment was characterized by the quadrupolar splitting of the 2H NMR water signal and by the measurement of 1H-15N residual dipolar couplings (RDC) in the archeal translation elongation factor 1beta. Protein dissolved in the polymer-stabilized medium orients quantitatively as in media without polyacrylamide. We show that the quadrupolar splitting and RDCs are zero in media in which the Pf1 phage particles are aligned at the magic angle. This allows measurement of J and dipolar couplings in a single sample.

摘要

一种新型聚合物稳定向列相液晶已被用于生物分子剩余偶极耦合的测量表征。丝状Pf1噬菌体被嵌入到固定颗粒取向的聚丙烯酰胺基质中。通过2H NMR水信号的四极分裂以及古细菌翻译延伸因子1β中1H-15N剩余偶极耦合（RDC）的测量来表征排列情况。溶解在聚合物稳定介质中的蛋白质与没有聚丙烯酰胺的介质中一样定量取向。我们表明，在Pf1噬菌体颗粒以魔角排列的介质中，四极分裂和RDC为零。这使得能够在单个样品中测量J和偶极耦合。