Geeves, Michael A. and Perreault-Micale, Cynthia and Coluccio, Lynne 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 currently available from this repository. You may be able to access a copy if URLs are provided)
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.
|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:||21 May 2014 07:39|
|Resource URI:||https://kar.kent.ac.uk/id/eprint/16132 (The current URI for this page, for reference purposes)|