MARCKS-like protein, a membrane protein identified for its expression in developing neural retina, plays a role in regulating retinal cell proliferation

Jing Zhao, Tomonori Izumi, Kazuto Nunomura, Shinya Satoh, Sumiko Watanabe

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Membrane proteins are expressed in a specific manner in developing tissues, and characterization of these proteins is valuable because it allows them to be used as cell surface markers. Furthermore, they are potentially important for the regulation of organogenesis because some may participate in signal transduction. In the present study, we used proteomics to examine the comprehensive protein expression profile of the membrane fraction in the embryonic and adult mouse retina. We purified the retinal membrane fraction by sucrose-density-gradient centrifugation and analysed total proteins using shotgun analysis on a nanoflow LC-MS/MS (liquid chromatography tandem MS) system. Approximately half of the 326 proteins from the adult retina and a quarter of the 310 proteins from the embryonic retina (day 17) appeared to be membrane-associated proteins. Among these, MLP [MARCKS (myristoylated alanine-rich C-kinase substrate)-like protein], which shares approx. 50% amino acid identity with MARCKS, was selected for further characterization. The mRNA and surface protein expression of MLP decreased as retinal development progressed. Overexpression of MLP by retrovirus-mediated gene transfer enhanced the proliferation of retinal progenitor cells without affecting differentiation or cell migration in a retinal expiant culture system. In contrast, MLP overexpression did not promote proliferation in fibroblasts (NIH 3T3 cells). Mutation analysis of MLP demonstrated that myristoylation was necessary to promote proliferation and that phosphorylation inhibited proliferation, indicating the functional importance of membrane localization.

Original languageEnglish
Pages (from-to)51-59
Number of pages9
JournalBiochemical Journal
Volume408
Issue number1
DOIs
Publication statusPublished - Nov 15 2007
Externally publishedYes

Keywords

  • Cell proliferation
  • Membrane protein
  • Myristoylated alanine-rich C-kinase substrate (MARCKS)
  • Neural retina
  • Protein kinase C (PKC)
  • Proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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