title: Controlled activation of protein rotational dynamics using smart hydrogel tethering.
abstract: 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 (13)C and (15)N, 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.
authors.affiliation: School of Biological Sciences, Washington State University Tri-Cities , Pullman, Washington 99164, United States.
authors.forename: Brenda M
authors.initials: BM
authors.lastname: Beech
authors.forename: Yijia
authors.initials: Y
authors.lastname: Xiong
authors.forename: Curt B
authors.initials: CB
authors.lastname: Boschek
authors.forename: Cheryl L
authors.initials: CL
authors.lastname: Baird
authors.forename: Diana J
authors.initials: DJ
authors.lastname: Bigelow
authors.forename: Kathleen
authors.initials: K
authors.lastname: McAteer
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Squier
mesh_terms: D001665:Binding Sites; D002147:Calmodulin / Q000737:chemistry*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry*; D058619:Maltose-Binding Proteins / Q000737:chemistry*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011092:Polyethylene Glycols / Q000737:chemistry*; D055550:Protein Stability; D024744:Skeletal Muscle Myosins / Q000737:chemistry*
chemical_list: D002147:Calmodulin; D055767:Immobilized Proteins; D058619:Maltose-Binding Proteins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D011092:Polyethylene Glycols; D024744:Skeletal Muscle Myosins