DOI: 10.1002/pro.5560050308. Interlobe communication in multiple calcium‐binding site mutants of <i>Drosophila</i> calmodulin

Interlobe communication in multiple calcium‐binding site mutants of Drosophila calmodulin

10.1002/pro.5560050308

Crossref journal-article

Wiley

Protein Science (311)

Abstract

AbstractWe have generated mutants of Drosophila calmodulin in which pairs of calcium‐binding sites are mutated so as to prevent calcium binding. In all sites, the mutation involves replacement of the —Z position glutamate residue with glutamine. Mutants inactivated in both N‐terminal sites (B12Q) or both C‐terminal sites (B34Q), and two mutants with one N‐ and one C‐terminal site inactivated (B13Q and B24Q) were generated. The quadruple mutant with all four sites mutated was also studied. UV‐difference spectroscopy and near‐UV CD were used to examine the influence of these mutations upon the single tyrosine (Tyr‐138) of the protein. These studies uncovered four situations in which Tyr‐138 in the C‐terminal lobe responds to a change in the calcium‐binding properties of the N‐terminal lobe. Further, they suggest that N‐terminal calcium‐binding events contribute strongly to the aberrant behavior of Tyr‐138 seen in mutants with a single functional C‐terminal calcium‐binding site. The data also indicate that loss of calcium binding at site 1 adjusts the aberrant conformation of Tyr‐138 produced by mutation of site 3 toward the wild‐type structure. However, activation studies for skeletal muscle myosin light chain kinase (SK‐MLCK) established that all of the multiple binding site mutants are poor activators of SK‐MLCK. Thus, globally, the calcium‐induced conformation of B13Q is not closer to wild type than that of either the site 1 or the site 3 mutant. The positioning of Tyr‐138 within the crystal structure of calmodulin suggests that effects of the N‐terminal lobe on this residue may be mediated via changes to the central linker region of the protein.

Bibliography

Mukherjea, P., Maune, J. F., & Beckingham, K. (1996). Interlobe communication in multiple calciumâbinding site mutants of Drosophila calmodulin. Protein Science, 5(3), 468â477. Portico.

Authors 3

Poushali Mukherjea (first)
John F. Maune (additional)
Kathy Beckingham (additional)

References 30 Referenced 32

10.1016/0022-2836(88)90608-0
10.1021/bi00138a005
10.1021/bi00255a006
{'key': 'e_1_2_1_5_1', 'first-page': '15259', 'article-title': 'Ultraviolet difference spectroscopy ‐ New techniques and applications', 'volume': '33', 'author': 'Donovan JW', 'year': '1973', 'journal-title': 'Biochemistry'} / Biochemistry / Ultraviolet difference spectroscopy ‐ New techniques and applications by Donovan JW (1973)
10.1016/B978-0-12-171406-2.50010-X
10.1016/S0021-9258(20)80699-1
10.1021/bi00403a011
10.1021/bi00102a013
10.1016/S0021-9258(19)44043-X / J Biol Chem / Carp muscle calcium‐binding protein by Kretsinger RH (1973)
10.1038/nsb0995-768
10.1016/S0021-9258(18)92938-8
10.1042/bj2380485
10.1111/j.1432-1033.1992.tb16879.x
10.1021/bi00149a006
10.1016/S0021-9258(18)42764-0 / J Biol Chem / Ca2+ binding and conformational change in two series of point mutations to the individual Ca2+ binding sites of calmodulin by Maune JF (1992)
MukherjeaP.1995.Studies of the roles of the individual calcium binding sites ofDrosophilacalmodulin in conformational change and target interaction [thesis]. Houston Texas: Rice University.
{'key': 'e_1_2_1_18_1', 'first-page': '555', 'article-title': 'Calcium binding site mutants of calmodulin adopt abnormal conformations in complexes with model target peptides', 'volume': '29', 'author': 'Mukherjea P', 'year': '1993', 'journal-title': 'Biochem Mol Biol Int'} / Biochem Mol Biol Int / Calcium binding site mutants of calmodulin adopt abnormal conformations in complexes with model target peptides by Mukherjea P (1993)
10.1080/00327488808062527
10.1021/bi00598a029
10.1016/S0021-9258(18)50605-0 / J Biol Chem / Specific perturbation by Ca2+ of tyrosyl residue 138 of calmodulin by Richman PG (1979)
10.1021/bi00542a031
10.1021/bi00345a002
10.1016/0022-2836(87)90025-8
10.1002/pro.5560010206
10.1002/prot.340030102
{'key': 'e_1_2_1_27_1', 'first-page': '12', 'article-title': 'Improved media for growing plasmid and cos‐mid clones', 'volume': '9', 'author': 'Tartof KD', 'year': '1987', 'journal-title': 'Bethesda Res Labs Focus'} / Bethesda Res Labs Focus / Improved media for growing plasmid and cos‐mid clones by Tartof KD (1987)
10.1016/S0021-9258(18)54647-0
10.1021/bi00128a020
10.1016/S0021-9258(18)54665-2 / J Biol Chem / Three amino acid substitutions in domain‐1 of calmodulin prevent the activation of chicken smooth muscle myosin light chain kinase by VanBerkum MRA (1991)
10.1038/nsb0995-758

Dates

Type	When
Created	15 years, 1 month ago (July 12, 2010, 1:15 a.m.)
Deposited	1 year, 10 months ago (Oct. 26, 2023, 11:43 p.m.)
Indexed	1 year, 7 months ago (Jan. 16, 2024, 6:21 a.m.)
Issued	29 years, 6 months ago (March 1, 1996)
Published	29 years, 6 months ago (March 1, 1996)
Published Online	16 years, 8 months ago (Dec. 31, 2008)
Published Print	29 years, 6 months ago (March 1, 1996)

Funders 2

NIH
Awards2
1. 003604-028
2. GM49155
Research/Advanced Technology Program of the Texas Higher Education Board, and Welch Foundation Grant
Awards1
1. C-1119

BibTeX

@article{Mukherjea_1996, title={Interlobe communication in multiple calcium‐binding site mutants of Drosophila calmodulin}, volume={5}, ISSN={1469-896X}, url={http://dx.doi.org/10.1002/pro.5560050308}, DOI={10.1002/pro.5560050308}, number={3}, journal={Protein Science}, publisher={Wiley}, author={Mukherjea, Poushali and Maune, John F. and Beckingham, Kathy}, year={1996}, month=mar, pages={468–477} }

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