Molecular cloning of glycoprotein antigens MGP57/53 recognized by monoclonal antibodies raised against bovine milk fat globule membrane.
A cDNA encoding 57 kDa and 53 kDa antigens (MGP57/53) recognized by monoclonal antibodies raised against bovine milk fat globule membrane (MFGM) (Biochim. Biophys. Acta 1199 (1994) 87-95) was cloned from lactating bovine mammary gland by a combination of reverse transcriptase-coupled polymerase chain reaction (RT-PCR) and 3'-rapid amplification of cDNA ends (3'-RACE). The deduced amino-acid sequence showed that mature MGP57/53 consists of 409 amino-acid residues and the calculated molecular weight and isoelectric point are 45,544 and 6.42, respectively. Computer analysis reveals that it has a significant similarity to mouse mammary epithelial cell surface protein, MFG-E8 and a human breast tumor-associated glycoprotein antigen, BA46-1. An N-terminal cysteine-rich domain and a C-terminal tandemly repeated sequence were highly conserved among them, but bovine MGP57/53 lacks 36 amino-acid residues containing a cluster of 5 prolines found in mouse MFG-E8. Northern blot analysis showed that the cDNA hybridized to about 2.0 kb mRNA of lactating bovine mammary gland. These results strongly support our previous report that the two MFGM antigens originate from a single gene and are isoforms with different N-linked sugar chains.
Department of Applied Biological Sciences, School of Agricultural Sciences, Nagoya University, Japan.
N
N
Aoki
M
M
Kishi
Y
Y
Taniguchi
T
T
Adachi
R
R
Nakamura
T
T
Matsuda
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002417:Cattle; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007774:Lactation; D008321:Mammary Glands, Animal / Q000378:metabolism*; D051379:Mice; D008894:Milk Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016415:Sequence Alignment
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D006023:Glycoproteins; D008894:Milk Proteins