GSBS Student Publications

Title

Essential role of Ca2+/calmodulin in Early Endosome Antigen-1 localization

UMMS Affiliation

Graduate School of Biomedical Sciences; Program in Molecular Medicine; Department of Physiology

Date

7-15-2003

Document Type

Article

Medical Subject Headings

1-Phosphatidylinositol 3-Kinase; Amino Acid Motifs; Animals; COS Cells; Calcium; Calmodulin; Cercopithecus aethiops; Endosomes; Liposomes; *Membrane Fusion; Membrane Proteins; Microscopy, Fluorescence; Mutation; Protein Structure, Quaternary; Recombinant Proteins; Sulfonamides; Vesicular Transport Proteins; rab5 GTP-Binding Proteins

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Ca2+ is an essential requirement in membrane fusion, acting through binding proteins such as calmodulin (CaM). Ca2+/CaM is required for early endosome fusion in vitro, however, the molecular basis for this requirement is unknown. An additional requirement for endosome fusion is the protein Early Endosome Antigen 1 (EEA1), and its recruitment to the endosome depends on phosphatidylinositol 3-phosphate [PI(3)P] and the Rab5 GTPase. Herein, we demonstrate that inhibition of Ca2+/CaM, by using either chemical inhibitors or specific antibodies directed to CaM, results in a profound inhibition of EEA1 binding to endosomal membranes both in live cells and in vitro. The concentration of Ca2+/CaM inhibitors required for a full dissociation of EEA1 from endosomal membranes had no effect on the activity of phosphatidylinositol 3-kinases or on endogenous levels of PI(3)P. However, the interaction of EEA1 with liposomes containing PI(3)P was decreased by Ca2+/CaM inhibitors. Thus, Ca2+/CaM seems to be required for the stable interaction of EEA1 with endosomal PI(3)P, perhaps by directly or indirectly stabilizing the quaternary organization of the C-terminal FYVE domain of EEA1. This requirement is likely to underlie at least in part the essential role of Ca2+/CaM in endosome fusion.

Rights and Permissions

Citation: Mol Biol Cell. 2003 Jul;14(7):2935-45. Epub 2003 Mar 20. Link to article on publisher's site

DOI of Published Version

10.1091/mbc.E02-09-0591

Related Resources

Link to article in PubMed

Journal Title

Molecular biology of the cell

PubMed ID

12857876