The isolation and characterization of plasma membrane from cultured cells I. The chemical composition of membrane isolated from uninfected and oncogenic RNA virus-converted chick embryo fibroblasts

1971 ◽  
Vol 249 (2) ◽  
pp. 419-434 ◽  
Author(s):  
James F. Perdue ◽  
Rolf Kletzien ◽  
Katherine Miller
Keyword(s):  
Plasma Membrane ◽  
Chemical Composition ◽  
Chick Embryo ◽  
Cultured Cells ◽  
Rna Virus ◽  
Isolation And Characterization ◽  
Embryo Fibroblasts

Isolation and characterization of a vinculin cDNA from chick embryo fibroblasts

1987 ◽  
Vol 15 (5) ◽  
pp. 794-796
Author(s):  
PETER JONES ◽  
GLYN J. PRICE ◽  
MATTHEW D. DAVISON ◽  
DAVID R. CRITCHLEY
Keyword(s):  
Chick Embryo ◽  
Isolation And Characterization ◽  
Embryo Fibroblasts

scholarly journals Isolation and characterization of a vinculin cDNA from chick-embryo fibroblasts

1987 ◽  
Vol 245 (2) ◽  
pp. 595-603 ◽  
Author(s):  
G J Price ◽  
P Jones ◽  
M D Davison ◽  
B Patel ◽  
I C Eperon ◽  
...  
Keyword(s):  
Chick Embryo ◽  
Fusion Protein ◽  
Tyrosine Kinases ◽  
Consensus Sequence ◽  
Coli Strain ◽  
Amino Acid Residues ◽  
Reading Frame ◽  
Internal Repeat ◽  
Isolation And Characterization ◽  
Embryo Fibroblasts

A chick-embryo fibroblast lambda gt11 cDNA library was screened with affinity-purified antibodies to chick gizzard vinculin. One recombinant was purified to homogeneity and the fusion protein expressed in Escherichia coli strain C600. The fusion protein was unstable, but polypeptides that reacted with vinculin antibodies, but not non-immune immunoglobulin, were detected by Western blotting. The recombinant contained a single EcoRI fragment of 2891 bp with a single open reading frame. The deduced protein sequence could be aligned with that of six CNBr-cleavage peptides and two tryptic peptides derived from chicken gizzard vinculin. AUG-247 has tentatively been identified as the initiation codon, as it is contained within the consensus sequence for initiation sites of higher eukaryotes. The cDNA lacks 3′ sequence and encodes 74% of the vinculin sequence, presuming the molecular mass of vinculin to be 130,000 Da. Analysis of the deduced sequence showed no homologies with other protein sequences, but it does display a triple internal repeat of 112 amino acid residues covering residues 259-589. The sequences surrounding the seven tyrosine residues in the available sequence were aligned with the tyrosine autophosphorylation consensus sequence found in protein tyrosine kinases. Tyr-822 showed a good match to this consensus, and may represent one of the two major sites of tyrosine phosphorylation by pp60v-sre. Northern blots showed that the 2.89 kb vinculin cDNA hybridized to one size of mRNA (approx. 7 kb) in chick-embryo fibroblasts, chick smooth muscle and chick skeletal muscle. Southern blots revealed multiple hybridizing bands in genomic DNA.

Sign in / Sign up

Export Citation Format

Share Document