• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Quantitative proteomic and lipidomic analysis of detergent-resistant membrane microdomain involved in signal transduction

Research Project

Project/Area Number 16570100
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionJUNTENDO UNIVERSITY

Principal Investigator

YANAGIDA Mitsuaki  Juntendo University, School of Medicine, Assistant Professor, 医学部, 講師 (80365569)

Co-Investigator(Kenkyū-buntansha) KAGA Naoko  Juntendo University, School of Medicine, Assistant, 医学部, 助手 (80338342)
IWABUCHI Kazuhisa  Juntendo University, School of Health Care and Nursing, Associate Professor, 医療看護学部, 助教授 (10184897)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2004: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywordsproteomics / lipidomics / microdomain / mass spectrometry / stable-isotope labeling / innate immunity / phagocytosis / 定量プロテオミクス / 質料分析
Research Abstract

Neutrophils, which share a major population of leukocytes, are responsible for killing of bacteria. Recent reports showed that plasma membrane microdomains (lipid rafts) of neutrophils mediated phagocytosis and that through phagocytosis, infectious microorganisms not only entered in the cells but also survived by use of microdomains. The membrane microdomains are sphingolipid- and cholesterol-rich domains distinct from other membrane surfaces and provide the characteristic regions where many specific proteins including signal-transducing molecules are clustered.
In order to elucidate the function of microdomains during the phagocytic process in neutrophils, we tried to obtain fundamental data of protein and lipid components related to this pathway. As a model of neutrophil, we used human promyelocytic leukemia cell line, HL-60, which could be differentiated into neutrophil-like cells by treatment with dimethylsulfoxide. We prepared the detergent-resistant membranes (DRMs) in HL-60 cells before and after differentiation, and performed proteomic analysis by nano-flow liquid chromatography-mass spectrometry (NanoLC-MS/MS).
NanoLC-MS/MS analysis identified 121 proteins, in which 41 and 25 proteins were mainly detected in untreated and DMSO-treated HL-60 DRMs, respectively. More than 30% of identified proteins were already identified membrane proteins. Otherwise, many G-proteins and cytoskeleton proteins were also detected. We thought that identified proteins that showed change during differentiation were candidate proteins involved in function of neutrophils. Quantitation with internal standard peptides labeled with stable 13C and 15N isotopes revealed that protein detection profile of transferrin receptor and flotillins corresponded with previously reported knowledge. We concluded that the quantitative proteomic analysis using stable isotopes were effective for elucidation of protein construction of microdomains.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report

URL: 

Published: 2004-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi