Kinetic analyses of a truncated mammalian myosin I suggest a novel isomerization event preceding nucleotide binding

Geeves, M.A. and Perreault-Micale, C and Coluccio, L.M. (2000) Kinetic analyses of a truncated mammalian myosin I suggest a novel isomerization event preceding nucleotide binding. Journal of Biological Chemistry, 275 (28). pp. 21624-21630. ISSN 0021-9258. (The full text of this publication is not available from this repository)

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Official URL
http://dx.doi.org/10.1074/jbc.M000342200

Abstract

MI1IQ is a complex of calmodulin and an epitope-tagged 85-kDa fragment representing the amino-terminal catalytic motor domain and the first of 6 calmodulin-binding IQ domains of the mammalian myosin I gene, rat myr-1 (130-kDa myosin I or MI130). We have determined the transient kinetic parameters that dictate the ATP hydrolysis cycle of mammalian myosin I by examining the properties of MI1IQ. Transient kinetics reveal that the affinity of MI1IQ for actin is 12 nM. The ATP-induced dissociation of actin-MI1IQ is biphasic. The fast phase is dependent upon [ATP], whereas the slow phase is not; both phases show a Ca2+ sensitivity. The fast phase is eliminated by the addition of ADP, 10 mu M being required for half-saturation of the effect in the presence of Ca2+ and 3 mu M ADP in the absence of Ca2+. The slow phase shares the same rate constant as ADP release (8 and 3 s(-1) in the presence and absence of Ca2+, respectively), but cannot be eliminated by decreasing [ADP]. We interpret these results to suggest that actin-myosin I exists in two forms in equilibrium, one of which is unable to bind nucleotide. These results also indicate that the absence of the COOH-terminal 5 calmodulin binding domains of myr-1 do not influence the kinetic properties of MI130 and that the Ca2+ sensitivity of the kinetics are in all likelihood due to Ca2+ binding to the first IQ domain.

Item Type: Article
Subjects: Q Science > QD Chemistry
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: O.O. Odanye
Date Deposited: 14 May 2009 10:43
Last Modified: 24 Apr 2012 13:48
Resource URI: http://kar.kent.ac.uk/id/eprint/16132 (The current URI for this page, for reference purposes)
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