Title

Molecular characterization of mitofilin (HMP), a mitochondria-associated protein with predicted coiled coil and intermembrane space targeting domains

UMMS Affiliation

Graduate School of Biomedical Sciences; Department of Cell Biology; Department of Biochemistry and Molecular Pharmacology

Date

9-1-1996

Document Type

Article

Medical Subject Headings

Actins; Amino Acid Sequence; Animals; Antibodies; Binding Sites; Cell Line; Cloning, Molecular; DNA, Complementary; Detergents; Humans; Mice; Microscopy, Fluorescence; Microscopy, Immunoelectron; Microtubules; Mitochondria; Mitochondrial Proteins; Molecular Sequence Data; Molecular Structure; Muscle Proteins; RNA, Messenger; Recombinant Fusion Proteins

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

We have identified and characterized a human protein of the mitochondria which we call mitofilin. Using monoclonal and polyclonal antibodies, we have isolated cDNA clones and characterized mitofilin biochemically. It appears as a 90 and 91 kDa doublet in western blots and is translated from a single 2.7 kb mRNA. Antibodies raised against cellular and bacterially-expressed protein given identical cytoplasmic immunofluorescence and immunoblot results. Mitofilin co-localizes with mitochondria in immunofluorescence experiments and co-purifies with mitochondria. Double label studies show co-localization only with mitochondria and not with Golgi or endoplasmic reticulum. Co-localization with mitochondria is retained when actin or tubulin are de-polymerized, and mitofilin is expressed in all human cell types tested. The cDNA encodes a polypeptide with a central alpha-helical region with predicted coiled coil domains flanked by globular amino and carboxy termini. Unlike coiled coil motor proteins, mitofilin is resistant to detergent extraction. The presence of mitochondrial targeting and stop-transfer sequences, along with the accessibility of mitofilin to limited proteolysis suggests that it resides predominantly in the intermembrane space, consistent with immuno-electron micrographs which show mitofilin mainly at the mitochondrial periphery. The cDNA sequence of mitofilin is identical to that recently reported by Icho et al. (1994; Gene 144, 301-306) for a mRNA preferentially expressed in heart muscle (HMP), consistent with the high levels of mitochondria in cardiac myocytes.

Rights and Permissions

Citation: J Cell Sci. 1996 Sep;109 ( Pt 9):2253-64.

Related Resources

Link to article in PubMed

PubMed ID

8886976