Allosteric factors in the calcium/calmodulin-responsive kinase II hub domain determine selectivity of GHB ligands for CaMKIIα
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F25%3A00635735" target="_blank" >RIV/61388963:_____/25:00635735 - isvavai.cz</a>
Result on the web
<a href="https://doi.org/10.1016/j.jbc.2025.108543" target="_blank" >https://doi.org/10.1016/j.jbc.2025.108543</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.jbc.2025.108543" target="_blank" >10.1016/j.jbc.2025.108543</a>
Alternative languages
Result language
angličtina
Original language name
Allosteric factors in the calcium/calmodulin-responsive kinase II hub domain determine selectivity of GHB ligands for CaMKIIα
Original language description
The Ca2+/CaM-dependent protein kinase II alpha (CaMKIIα) is a highly important synaptic protein, which comprises a unique holoenzyme structure organized via the central hub domain. Recently, a distinct binding pocket in the CaMKIIα hub domain was identified for the endogenous neuromodulator γ-hydroxybutyric acid (GHB) and related synthetic analogs. Intriguingly, of the four native CaMKII isozymes, only CaMKIIα accommodates GHB ligands. Key interacting residues in CaMKIIα were revealed, but their involvement in selectivity toward the alpha variant of CaMKII has remained unresolved. Aimed at elucidating the molecular determinants for this selectivity, we here conducted binding studies to CaMKII-HEK whole-cell homogenates using two different in-house–developed GHB-related radioligands, 3-hydroxycyclopent-1-enecarboxylic acid ([3H]HOCPCA) and [3H]O-5-hydroxydiclofenac, in combination with site-directed mutagenesis. Binding to CaMKIIα with the smaller type radioligand [3H]HOCPCA validated key involvement of the four known residues (His395, Arg433, Arg453, and Arg469), but also revealed a role for the upper hub flexible loop containing the CaMKIIα-specific residue Trp403 (Leu in all other CaMKII isozymes) previously suggested to be involved in holoenzyme stability. Insertion of the corresponding residues (L467W/C533R) into CaMKIIβ failed to induce [3H]HOCPCA binding. However, with the larger type radioligand, [3H]O-5-hydroxydiclofenac, specific binding in CaMKIIβ (L467W/C533R) was achieved. Thus, the study confirms involvement of central binding residues and identifies the CaMKIIα flexible pocket loop as a distantly located allosteric factor in determining selectivity of GHB analogs for CaMKIIα. It sheds light on a remarkable interplay of the entire hub cavity for accommodation of ligands and corroborates GHB analogs as CaMKIIα-selective.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10406 - Analytical chemistry
Result continuities
Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2025
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Biological Chemistry
ISSN
0021-9258
e-ISSN
1083-351X
Volume of the periodical
301
Issue of the periodical within the volume
6
Country of publishing house
NL - THE KINGDOM OF THE NETHERLANDS
Number of pages
10
Pages from-to
108543
UT code for WoS article
001498943500001
EID of the result in the Scopus database
2-s2.0-105005486026