Neurogranin Modulates the Rate of Association Between Calmodulin and Target Peptides

Authors

John A Putkey
Laurel Hoffman
Vladimir Berka
Xu Wang

Language

English

Publication Date

6-18-2024

Journal

Biophysical Journal

DOI

10.1016/j.bpj.2024.05.010

PMID

38751114

PMCID

PMC11213993

PubMedCentral® Posted Date

5-14-2024

PubMedCentral® Full Text Version

Post-print

Abstract

The best-known mode of action of calmodulin (CaM) is binding of Ca2+ to its N- and C-domains, followed by binding to target proteins. An underappreciated facet of this process is that CaM is typically bound to proteins at basal levels of free Ca2+, including the small, intrinsically disordered, neuronal IQ-motif proteins called PEP-19 and neurogranin (Ng). PEP-19 and Ng would not be effective competitive inhibitors of high-affinity Ca2+-dependent CaM targets at equilibrium because they bind to CaM with relatively low affinity, but they could influence the time course of CaM signaling by affecting the rate of association of CaM with high-affinity Ca2+-dependent targets. This mode of regulation may be domain specific because PEP-19 binds to the C-domain of CaM, whereas Ng binds to both N- and C-domains. In this report, we used a model CaM binding peptide (CKIIp) to characterize the preferred pathway of complex formation with Ca2+-CaM at low levels of free Ca2+ (0.25–1.5 μM), and how PEP-19 and Ng affect this process. We show that the dominant encounter complex involves association of CKIIp with the N-domain of CaM, even though the C-domain has a greater affinity for Ca2+. We also show that Ng greatly decreases the rate of association of Ca2+-CaM with CKIIp due to the relatively slow dissociation of Ng from CaM, and to interactions between the Gly-rich C-terminal region of Ng with the N-domain of CaM, which inhibits formation of the preferred encounter complex with CKIIp. These results provide the general mechanistic paradigms that binding CaM to targets can be driven by its N-domain, and that low-affinity regulators of CaM signaling have the potential to influence the rate of activation of high-affinity CaM targets and potentially affect the distribution of limited CaM among multiple targets during Ca2+ oscillations.

Keywords

Calmodulin, Neurogranin, Protein Binding, Calcium, Peptides, Protein Domains, Kinetics, Amino Acid Sequence, Animals

Published Open-Access

yes

Recommended Citation

Putkey, John A; Hoffman, Laurel; Berka, Vladimir; et al., "Neurogranin Modulates the Rate of Association Between Calmodulin and Target Peptides" (2024). Faculty, Staff and Student Publications. 6454.
https://digitalcommons.library.tmc.edu/uthgsbs_docs/6454