tailieunhanh - Báo cáo khoa học: Contributions to catalysis and potential interactions of the three catalytic domains in a contiguous trimeric creatine kinase

Three separate creatine kinase (CK) isoform families exist in animals. Two of these (cytoplasmic and mitochondrial) are obligate oligomers. A third, flagellar, is monomeric but contains the residues for three complete CK domains. | ễFEBS Journal Contributions to catalysis and potential interactions of the three catalytic domains in a contiguous trimeric creatine kinase Gregg G. Hoffman1 Omar Davulcu2 Sona Sona1 and W. Ross Ellington1 3 1 Department of BiologicalScience Florida State University Tallahassee FL USA 2 Department of Chemistry and Biochemistry Florida State University Tallahassee FL USA 3 Institute of Molecular Biophysics Florida State University Tallahassee FL USA Keywords contiguous trimer cooperativity domain interaction flagellar creatine kinase kinetics Correspondence W. R. Ellington Institute of Molecular Biophysics Florida State University Tallahassee FL USA Fax 1 850 644 0481 Tel 1 850 644 5406 E-mail elling@ Received 23 July 2007 revised 26 November 2007 accepted 10 December 2007 doi Three separate creatine kinase CK isoform families exist in animals. Two of these cytoplasmic and mitochondrial are obligate oligomers. A third flagellar is monomeric but contains the residues for three complete CK domains. It is not known whether the active sites in each of the contiguous flagellar domains are catalytically competent and if so whether they are capable of acting independently. Here we have utilized site-directed mutagenesis to selectively disable individual active sites and all possible combinations thereof. Kinetic studies showed that these mutations had minimal impact on substrate binding and synergism. Interestingly the active sites were not catalytically equivalent and were in fact interdependent a phenomenon that has previously been reported only in the oligomeric CK isoforms. Creatine kinase CK plays a central role in energy homeostasis in cells that display high or variable rates of ATP utilization such as neurons muscle fibers transport epithelia and spermatozoa 1 . The physiological roles of the CK reaction are greatly facilitated by the presence of three nuclear gene families each targeted to and localized in specific .

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