Characterization through cDNA cloning of galactoprotein b3 (Gap b3), a cell surface membrane glycoprotein showing enhanced expression on oncogenic transformation. Identification of Gap b3 as a member of the integrin superfamily.
Galactoprotein b (Gap b) is a group of glycoproteins of fibroblast cell membranes showing enhanced expression in association with oncogenic transformation (Carter, W.G., and Hakomori, S. (1977) Biochem. Biophys. Res. Commun. 76, 299-308). We purified Gap b3 (one of the Gap b group) to homogeneity from hamster fibroblast NIL cells transformed with polyoma virus (NILpy). Partial amino acid sequence was determined, and the cDNA clones encoding this protein were isolated by oligodeoxynucleotide hybridization. The 5.0-kilobase pair cDNA encodes a protein of 1051 amino acid residues containing a signal peptide (32 residues), a large extracellular domain (959 residues), a transmembrane domain (28 residues), and a short cytoplasmic domain (32 residues). The overall structure of Gap b3 is similar to that of the alpha subunit of so-called integrin receptors in the following respects: (i) presence of metal-binding sequences; (ii) location of a transmembrane domain near C terminus; (iii) matched positions of most of the cysteine residues. This, together with extensive amino acid sequence homology, strongly suggests that Gap b3 is a member of the integrin family. Gap b3 consists of two polypeptide chains (Mr = 110,000 and 30,000), which seem to be proteolytic cleavage products connected by disulfide bonds from a precursor protein. Southern hybridization shows the presence of a single copy gene in mice and humans as well. Northern hybridization analysis shows that the enhanced expression of Gap b3 in transformed cells is due to increased transcription of its message.
Biomembrane Institute, Seattle, Washington.
T
T
Tsuji
F
F
Yamamoto
Y
Y
Miura
K
K
Takio
K
K
Titani
S
S
Pawar
T
T
Osawa
S
S
Hakomori
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002471:Cell Transformation, Neoplastic; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D015723:Gene Library; D006023:Glycoproteins / Q000235:genetics*; D015151:Immunoblotting; D039201:Integrin alpha3beta1; D016023:Integrins / Q000235:genetics* / Q000302:isolation & purification; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D005810:Multigene Family; D015345:Oligonucleotide Probes; D011120:Polyomavirus / Q000235:genetics; D011487:Protein Conformation; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D006023:Glycoproteins; D039201:Integrin alpha3beta1; D016023:Integrins; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D015345:Oligonucleotide Probes; C017336:galactoproteins