Controlled Activation of Protein Rotational Dynamics Using Smart Hydrogel Tethering

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• 作者：Brenda M. Beech ; Yijia Xiong ; Curt B. Boschek ; Cheryl L. Baird ; Diana J. Bigelow ; Kathleen McAteer ; Thomas C. Squier
• 刊名：Journal of the American Chemical Society
• 出版年：2014
• 出版时间：September 24, 2014
• 年：2014
• 卷：136
• 期：38
• 页码：13134-13137
• 全文大小：269K
• ISSN：1520-5126

文摘

Stimulus-responsive hydrogel materials that stabilize and control protein dynamics have the potential to enable a range of applications that take advantage of the inherent specificity and catalytic efficiencies of proteins. Here we describe the modular construction of a hydrogel using an engineered calmodulin (CaM) within a poly(ethylene glycol) (PEG) matrix that involves the reversible tethering of proteins through an engineered CaM-binding sequence. For these measurements, maltose binding protein (MBP) was isotopically labeled with ¹³C and ¹⁵N, permitting dynamic structural measurements using TROSY-HSQC NMR spectroscopy. The protein dynamics is suppressed upon initial formation of hydrogels, with a concomitant increase in protein stability. Relaxation of the hydrogel matrix following transient heating results in enhanced protein dynamics and resolution of substrate-induced large-amplitude domain rearrangements.