Molecular Cloning of Human 14-3-3 cDNA with CD3 Zeta Peptide Containing Phosphosering Tag.
- Author:
Jae Seung KANG
- Publication Type:Original Article
- Keywords:
Cdna cloning;
ITAM
- MeSH:
14-3-3 Proteins;
Animals;
B-Lymphocytes;
Base Sequence;
Blotting, Western;
Clone Cells;
Cloning, Molecular*;
DNA, Complementary*;
HeLa Cells;
Humans*;
Mass Screening;
Peptides;
Phosphoserine;
Protein Isoforms;
Rats
- From:Korean Journal of Immunology
1998;20(3):317-323
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
14-3-3 proteins are highly conserved proteins of about 29 kDa and have a minimum of seven isoforms. 14- 3-3 proteins interact with many signalling proteins by the recognition of phosphoserine. For the identification of proteins which react with ITAM (immunoreceptor tyrosine-based activation motif) of CD3 zeta chain, labeled synthetic peptides representing the CD3 zeta chain structual motifs (ITAMs) with a tag of PKC substrate sequence were used for western blotting. One major protein band of approximately 29 kDa was identified in lysate of Jurkat T cell, B cells and HeLa cells. Screening of lamda gt 11 library derived from HeLa cell gave two clones of 14-3-3 protein cDNA. Inspection of their nucleotide sequences identified these two full length cDNA clones as the 29 kDa human homologue of rat 14-3-3 gamma and the human 14-3-3 zeta protein. The human 14-3-3 gamma isoform also showed high homology with other species in amino acid and nucleotide sequence. Although 14-3-3 proteins are phosphoserine-binding proteins, there may be another way of interaction between ITAMs of CD3 and 14-3-3 proteins.
