Molecular dynamics simulations integrating kinetics for Pb2+-induced arginine kinase inactivation and aggregation

Si, Yue-Xiu; Lee, Jinhyuk; Cai, Yan; Yin, Sheng; Yang, Jun‐Mo; Park, Yong‐Doo; Qian, Guo‐Ying

doi:10.1016/j.procbio.2015.02.008

Cited by 4 publications

(1 citation statement)

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“…Spectra were processed using NMRPipe ( Delaglio et al, 1995 ). The cross-peak intensities were obtained using Sparky ( Si et al, 2015 ). Sequence-specific assignments of the 1 H N and 15 N resonances for apo C1B-C2 were obtained using 2 H-decoupled 3D HN(CA)CB, HNCA(CB), HN(COCA)CB, and HN(CO)CA ( Yamazaki et al, 1994 ) experiments on a [U- 13 C, 15 N; 55%- 2 H] C1B-C2 sample.…”

Section: Methodsmentioning

confidence: 99%

Reactivity of Thiol-Rich Zn Sites in Diacylglycerol-Sensing PKC C1 Domain Probed by NMR Spectroscopy

Cole

Igumenova

2021

Front. Mol. Biosci.

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Conserved homology 1 (C1) domains are peripheral zinc finger domains that are responsible for recruiting their host signaling proteins, including Protein Kinase C (PKC) isoenzymes, to diacylglycerol-containing lipid membranes. In this work, we investigated the reactivity of the C1 structural zinc sites, using the cysteine-rich C1B regulatory region of the PKCα isoform as a paradigm. The choice of Cd2+ as a probe was prompted by previous findings that xenobiotic metal ions modulate PKC activity. Using solution NMR and UV-vis spectroscopy, we found that Cd2+ spontaneously replaced Zn2+ in both structural sites of the C1B domain, with the formation of all-Cd and mixed Zn/Cd protein species. The Cd2+ substitution for Zn2+ preserved the C1B fold and function, as probed by its ability to interact with a potent tumor-promoting agent. Both Cys3His metal-ion sites of C1B have higher affinity to Cd2+ than Zn2+, but are thermodynamically and kinetically inequivalent with respect to the metal ion replacement, despite the identical coordination spheres. We find that even in the presence of the oxygen-rich sites presented by the neighboring peripheral membrane-binding C2 domain, the thiol-rich sites can successfully compete for the available Cd2+. Our results indicate that Cd2+ can target the entire membrane-binding regulatory region of PKCs, and that the competition between the thiol- and oxygen-rich sites will likely determine the activation pattern of PKCs.

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Section: Methodsmentioning

confidence: 99%