Myristoylated and nonmyristoylated pools of sea urchin sperm flagellar creatine kinase exist side-by-side: myristoylation is necessary for efficient lipid association

Biochemistry
A F QuestR A McIlhinney

Abstract

In sperm of the sea urchin Strongylocentrotus purpuratus, a functional phosphocreatine shuttle, that requires the existence of mitochondrial and cytosolic creatine kinase (CK) isoforms in distinct locations, is essential for sperm motility. S. purpuratus sperm have an unusually large, 145 kDa CK isoform, present exclusively in the sperm tail (TCK), that is enriched in flagellum membrane preparations. Purified TCK contains two very similar proteins, designated TCKI and TCKII, of which only TCKII associates readily with liposomes and detergent micelles in vitro. Here we demonstrate by gas chromatography/mass spectrometry combined with selective ion monitoring that ions diagnostic for the presence of myristoylglycine in proteins are found in TCKII, but not TCKI. By contrast, TCKI, but not TCKII, served in vitro as a substrate for recombinant, polyhistidine-tagged N-myristoyltransferase and was myristoylated to high stoichiometries (0.58 +/- 0.14 pmol of myristate/pmol of TCK), in the presence of myristoyl-CoA, on glycine in amide linkage. In vitro myristoylated TCKI associated with phosphatidylcholine (PC)/phosphatidylserine (PS) (75:25) liposomes and Triton X-100 detergent micelles in gel filtration assays and with PC/PS liposome...Continue Reading

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Citations

Oct 16, 2004·Journal of Molecular Evolution·Tomohiko SuzukiW Ross Ellington
Mar 31, 2011·Amino Acids·Theo WallimannUwe Schlattner
Jul 9, 1999·Progress in Lipid Research·J C DeMarR E Anderson
Feb 22, 2001·Annual Review of Physiology·W R Ellington
Jan 24, 2007·Traffic·Søren T ChristensenPeter Satir
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Dec 6, 2006·The Journal of Biological Chemistry·Masashi KinukawaVictor D Vacquier
Mar 17, 1999·Analytical Biochemistry·J L KerwinJ R Yates

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