Urea-induced denaturation of human calcium/calmodulin-dependent protein kinase IV: a combined spectroscopic and MD simulation studies

Journal of Biomolecular Structure & Dynamics
Huma NazMd Imtaiyaz Hassan

Abstract

Calcium/calmodulin-dependent protein kinase IV (CaMKIV) is a multifunctional enzyme which belongs to the Ser/Thr kinase family. CaMKIV plays important role in varieties of biological processes such as gene expression regulation, memory consolidation, bone growth, T-cell maturation, sperm motility, regulation of microtubule dynamics, cell-cycle progression, and apoptosis. To measure stability parameters, urea-induced denaturation of CaMKIV was carried out at pH 7.4 and 25°C, using three different probes, namely far-UV CD, near-UV absorption, and tryptophan fluorescence. A coincidence of normalized denaturation curves of these optical properties suggests that urea-induced denaturation is a two-state process. Analysis of these denaturation curves gave values of 4.20 ± 0.12 kcal mol(-1), 2.95 ± 0.15 M, and 1.42 ± 0.06 kcal mol(-1) M(-1) for [Formula: see text] (Gibbs free energy change (ΔGD) in the absence of urea), Cm (molar urea concentration ([urea]) at the midpoint of the denaturation curve), and m (=∂ΔGD/∂[urea]), respectively. All these experimental observations have been fully supported by 30 ns molecular dynamics simulation studies.

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Citations

Apr 20, 2016·Journal of Biomolecular Structure & Dynamics·Amresh PrakashMd Imtaiyaz Hassan
May 14, 2016·Journal of Biomolecular Structure & Dynamics·Parvez KhanMd Imtaiyaz Hassan
Dec 23, 2017·Journal of Biomolecular Structure & Dynamics·Sunayana Begum SyedMd Imtaiyaz Hassan
May 29, 2019·Journal of Cellular Biochemistry·Ahmad Abu Turab NaqviImtaiyaz Hassan
Mar 7, 2020·Journal of Biomolecular Structure & Dynamics·Ahmad Abu Turab NaqviMd Imtaiyaz Hassan
Jul 15, 2020·Journal of Biomolecular Structure & Dynamics·Khalid A AlamryMd Imtaiyaz Hassan
Jul 30, 2020·Oxidative Medicine and Cellular Longevity·Preeti GuptaMd Imtaiyaz Hassan
Sep 10, 2019·Journal of Biomolecular Structure & Dynamics·Saman FatimaMd Imtaiyaz Hassan
Dec 19, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Taj MohammadImtaiyaz Hassan
Oct 16, 2020·Cell Biochemistry and Biophysics·Kehinde F OmolabiMahmoud E S Soliman

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Methods Mentioned

BETA
glycosylation
gel filtration
electrophoresis
circular dichroism
Fluorescence
protein folding

Software Mentioned

CHARMM
Discovery Studio
GROMACS
Dichroweb K2D server

Related Concepts

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