RRC ID 1305
Author Oikawa K, Kimura S, Aoki N, Atsuta Y, Takiyama Y, Nagato T, Yanai M, Kobayashi H, Sato K, Sasajima T, Tateno M.
Title Neuronal calcium sensor protein visinin-like protein-3 interacts with microsomal cytochrome b5 in a Ca2+-dependent manner.
Journal J Biol Chem
Abstract Visinin-like protein-3, which is one of the neuronal calcium sensors, has been shown to be mainly expressed in cerebellar Purkinje cells, but cellular function of this protein has not yet been elucidated. We examined the tissue distribution of murine visinin-like protein-3 transcripts using real-time reverse transcription-PCR. Visinin-like protein-3 mRNA was found to be expressed in peripheral tissues. Particularly, the expression of the transcript in the thymus was significantly higher than in other peripheral tissues. In addition, B6RVTC1 thymoma cells robustly expressed visinin-like protein-3 mRNA. To identify a target protein of visinin-like protein-3, we performed a pull-down experiment using glutathione S-transferase-tagged visinin-like protein-3 and two-dimensional electrophoresis. We demonstrated that microsomal cytochrome b(5) was a Ca(2+)-dependent binding partner of visinin-like protein-3. In a co-immunoprecipitation experiment, it was observed that hippocalcin, as well as visinin-like protein-3, could interact with cytochrome b(5). Furthermore, we confirmed that the sequence Val(114)-Tyr(127) at the C-terminal tail of cytochrome b(5) is the minimal structural requirement for binding to visinin-like protein-3. In addition, the loop His(19)-His(25) at the N terminus of visinin-like protein-3 is essential for binding to cytochrome b(5). Microsomal cytochrome b(5) was also shown to be a potential activator of cytochrome P450. The present findings raise the possibility that visinin-like protein-3 may link Ca(2+) signaling to the machinery of microsomal monooxygenase complex composed of cytochrome b(5), cytochrome P450, and some reductases. This report provides the first evidence of an interaction between visinin-like protein-3 and microsomal cytochrome b(5).
Volume 279(15)
Pages 15142-52
Published 2004-4-9
DOI 10.1074/jbc.M312766200
PII M312766200
PMID 14739275
MeSH Amino Acid Sequence Animals Blotting, Western Calcium / metabolism* Calcium-Binding Proteins / chemistry* Calcium-Binding Proteins / metabolism* Cattle Cell Line Cell Line, Tumor Cloning, Molecular Cytochrome P-450 Enzyme System / metabolism Cytochromes b5 / chemistry Cytochromes b5 / metabolism* Electrophoresis, Gel, Two-Dimensional Genetic Vectors Glutathione Transferase / metabolism Humans Mice Mice, Inbred C57BL Microsomes / metabolism Models, Biological Models, Molecular Molecular Sequence Data Mutation Nerve Tissue Proteins / chemistry* Nerve Tissue Proteins / metabolism* Neurocalcin Neurons / metabolism* Precipitin Tests Protein Binding Protein Structure, Tertiary RNA, Messenger / metabolism Receptors, Calcium-Sensing / metabolism* Recombinant Fusion Proteins / metabolism Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid Transcription, Genetic Transfection Tyrosine / chemistry Valine / chemistry
IF 4.106
Times Cited 17
Human and Animal Cells M1