Abstract
A regulatory region involved in both autoinhibition and calmodulin (CaM) binding has previously been identified in the multifunctional Ca2+/CaM-dependent protein kinase (CaM kinase II). We have tested the role of various segments of the regulatory region in autoinhibition by the analysis of a series of truncation, substitution, and deletion mutants of the CaM kinase II α subunit (CaM kinase IIα). Unexpectedly, the sequence Lys-Lys-Phe-Asn at positions 291-294, adjacent to the CaM binding domain, was found to be sufficient to maintain an inhibited state in a truncated form of the kinase. However, these residues are not essential in the context of the full-length protein, indicating the importance of additional residues from the overlapping CaM binding domain. We propose here a molecular model for CaM kinase IIα based on the three-dimensional structure of the cAPK-PKI-(5-24) (protein kinase inhibitor fragment) complex. It is predicted from this model that autoinhibition is of the pseudosubstrate variety and that autophosphorylation of Thr-286 could occur by an intersubunit reaction in the holoenzyme complex.
| Original language | English |
|---|---|
| Pages (from-to) | 12127-12131 |
| Number of pages | 5 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 89 |
| Issue number | 24 |
| State | Published - Dec 1 1992 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Genetics
- General
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Regulation of intrasteric inhibition of the multifunctional calcium/calmodulin-dependent protein kinase. / Cruzalegui, Francisco H.; Kapiloff, Michael S; Morfin, John Paul; Kemp, Bruce E.; Rosenfeld, Michael G.; Means, Anthony R.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 24, 01.12.1992, p. 12127-12131.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Regulation of intrasteric inhibition of the multifunctional calcium/calmodulin-dependent protein kinase
AU - Cruzalegui, Francisco H.
AU - Kapiloff, Michael S
AU - Morfin, John Paul
AU - Kemp, Bruce E.
AU - Rosenfeld, Michael G.
AU - Means, Anthony R.
PY - 1992/12/1
Y1 - 1992/12/1
N2 - A regulatory region involved in both autoinhibition and calmodulin (CaM) binding has previously been identified in the multifunctional Ca2+/CaM-dependent protein kinase (CaM kinase II). We have tested the role of various segments of the regulatory region in autoinhibition by the analysis of a series of truncation, substitution, and deletion mutants of the CaM kinase II α subunit (CaM kinase IIα). Unexpectedly, the sequence Lys-Lys-Phe-Asn at positions 291-294, adjacent to the CaM binding domain, was found to be sufficient to maintain an inhibited state in a truncated form of the kinase. However, these residues are not essential in the context of the full-length protein, indicating the importance of additional residues from the overlapping CaM binding domain. We propose here a molecular model for CaM kinase IIα based on the three-dimensional structure of the cAPK-PKI-(5-24) (protein kinase inhibitor fragment) complex. It is predicted from this model that autoinhibition is of the pseudosubstrate variety and that autophosphorylation of Thr-286 could occur by an intersubunit reaction in the holoenzyme complex.
AB - A regulatory region involved in both autoinhibition and calmodulin (CaM) binding has previously been identified in the multifunctional Ca2+/CaM-dependent protein kinase (CaM kinase II). We have tested the role of various segments of the regulatory region in autoinhibition by the analysis of a series of truncation, substitution, and deletion mutants of the CaM kinase II α subunit (CaM kinase IIα). Unexpectedly, the sequence Lys-Lys-Phe-Asn at positions 291-294, adjacent to the CaM binding domain, was found to be sufficient to maintain an inhibited state in a truncated form of the kinase. However, these residues are not essential in the context of the full-length protein, indicating the importance of additional residues from the overlapping CaM binding domain. We propose here a molecular model for CaM kinase IIα based on the three-dimensional structure of the cAPK-PKI-(5-24) (protein kinase inhibitor fragment) complex. It is predicted from this model that autoinhibition is of the pseudosubstrate variety and that autophosphorylation of Thr-286 could occur by an intersubunit reaction in the holoenzyme complex.
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M3 - Article
C2 - 1334558
AN - SCOPUS:0027070736
VL - 89
SP - 12127
EP - 12131
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 24
ER -