Cloning of a SH3 Domain-Containing Proline-Rich Protein, p85SPR, and Its Localization in Focal Adhesion

A mouse thymus cDNA expression library was screened with monoclonal antibody (mAb), B16-5 which binds to common epitope in SH3 domains of phospholipase C-γ 1 (PLC-γ 1) and Nck. We have determined the complete nucleotide sequence of one of several positive clones. The 4,172 bp cDNA clone (GenBank Acc...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 1997-06, Vol.235 (3), p.794-798
Main Authors: Oh, Woo Kyun, Yoo, Jae Cheal, Jo, Donggyu, Song, Young Hwa, Kim, Moon Gyo, Park, Dongeun
Format: Article
Language:English
Subjects:
Adaptor Proteins, Signal Transducing
Amino Acid Sequence
Animals
Antibodies, Monoclonal
Brain - metabolism
Cell Adhesion
Cell Line
Cloning, Molecular
DNA, Complementary
Epitopes - analysis
Gene Library
Isoenzymes - chemistry
Male
Mice
Mice, Inbred Strains
Molecular Sequence Data
Oncogene Proteins - chemistry
Peptide Biosynthesis
Peptides - chemistry
Phospholipase C gamma
Proline-Rich Protein Domains
Rats
Recombinant Proteins - biosynthesis
Recombinant Proteins - chemistry
Sequence Alignment
Sequence Homology, Amino Acid
Spodoptera
src Homology Domains
Testis - metabolism
Thymus Gland - enzymology
Transfection
Type C Phospholipases - chemistry
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Summary:A mouse thymus cDNA expression library was screened with monoclonal antibody (mAb), B16-5 which binds to common epitope in SH3 domains of phospholipase C-γ 1 (PLC-γ 1) and Nck. We have determined the complete nucleotide sequence of one of several positive clones. The 4,172 bp cDNA clone (GenBank Accession No. U96634) encodes a SH3 domain-containing protein of 646 amino acids. Besides the SH3 domain, the predicted protein has a proline-rich region, nuclear localization signals, and leucine zipper motifs. The expressed protein in Sf9 insect cell exhibits a polypeptide of 85 kDa on SDS-PAGE. The protein is widely distributed in rat tissue with an especially high level of expression in brain and testis. Interestingly, the specific antibodies detected four related proteins of different size (75, 85, 90 and 105 kDa) in brain. In A431 cell, p85SPR is enriched at focal adhesion points indicating that the protein may interact with protein(s) in focal complexes.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1997.6875