Localization and function of enzymes: role of arginine kinase in cilia of Tetrahymena pyriformis

2015 Fiscal Year Final Research Report

• Project/Area Number: 24570087
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Animal physiology/Animal behavior
• Research Institution: Kochi University
• Principal Investigator: Suzuki Tomohiko 高知大学, 自然科学系, 教授 (60145109)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 酵素の局在 / アルギニンキナーゼ / 繊毛運動

Outline of Final Research Achievements

Tetrahymena pyriformis contains two arginine kinases (AKs), a typical 40-kDa AK (AK1) and a two-domain 80-kDa enzyme (AK2). Interestingly, the amino acid sequence of T. pyriformis AK1, had a distinct myristoylation signal sequence at the N-terminus. In this work, we isolated and cloned the N-myristoyltransferase (NMT) cDNA from T. pyriformis. Then the NMT assay were performed, using the recombinant NMT and the N-terminal peptide GCSNSRSG of T. pyriformis AK1 or the peptide CSNSRSG which lacks N-terminal G of the former peptide. The use of the former peptide gave a typical Michaelis-Menten curve, while that for the latter gave no activity. A sufficient amount of modified AK1 enzyme was synthesized using an insect cell-free protein synthesis system. Then peptide mass fingerprinting (PMF) analyses were performed. The target N-myristoylated peptide was clearly observed with an experimental mass (m/z-H+) of 832.4747, indicating that the synthesized T. pyriformis AK1 is myristoylated.

Free Research Field

比較生化学