2017 Fiscal Year Final Research Report
Elucidation of substrate recognition of choline lysoplasmalogen-specific phospholipase D
| Project/Area Number |
15K05557
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Research Field |
Bio-related chemistry
|
|---|
| Research Institution | Fukushima University |
|---|
Principal Investigator |
|
|---|
| Co-Investigator(Renkei-kenkyūsha) |
MURAYAMA Kazutaka 東北大学, 大学院医工学研究科, 准教授 (40400452)
|
|---|
| Project Period (FY) |
2015-04-01 – 2018-03-31
|
| Keywords | Phospholipase D / ホスホリパーゼD（PLD） / Lysoplasmalogen-PLD / Choline lysoplasmalogen / Lyso-PAF（リゾ型血小板因子） / 基質認識メカニズム / 基質特異性改変 / 構造活性（機能）相関 |
|---|
| Outline of Final Research Achievements |
Choline lysoplasmalogen-specific phospholipase D (LyPlsCho-PLD) is an enzyme that catalyzes the hydrolytic cleavage of the phosphoester bond of LyPls, releasing choline and lysoplasmenic acid. LyPlsCho-PLD exhibited weak activity toward LysoPAF. Our goal is to elucidate the substrate recognition mechanism of LyPlsCho-PLD. Based on the homology-modeled structure of LyPls-PLD and the docking model with substrate analog, amino acid residues predicted to interact with the head group and sn-1-alkenyl ether chain were substituted by site-directed mutagenesis. The princial conclusions of the study by mutation analysis and kinetics were as follows: 1) M71, N173 and F211 of LyPlsCho-PLD may be involved in the recognition of sn-1 alkenyl ether chain of LyPlsCho. F211 and W282 may take part in substrate binding and reaction products release (=k+2). 2) F211A, I, L, and C variants remarkably altered in the substrate preference (LysoPAF >> LyPlsCho) while maintaining the high activity.
|
| Free Research Field |
酵素工学
|
