Purification of MUC1 from bovine milk-fat globules and characterization of a corresponding full-length cDNA clone.
The highly glycosylated protein MUC1 was purified from bovine milk-fat globule membranes by a procedure involving detergent extraction, ion-exchange chromatography and reverse-phase chromatography. The identity of the purified mucin protein was confirmed by N-terminal sequencing and partial amino acid sequences obtained by peptide mapping. The complete amino acid sequence of MUC1 was determined by cloning and sequencing the corresponding bovine mammary gland cDNA, which was shown to encode a protein of 580 amino acid residues comprising a cleavable signal peptide of 22 residues. The deduced amino acid sequence demonstrated structural features characteristic for mucins, including an extracellular tandem repeat region with 11 partially conserved repeats (20 amino acids each), a membrane-proximal SEA module, a transmembrane domain, and a cytoplasmic C-terminal region. Monosaccharide composition determinations suggested significant structural differences between O-linked glycans of MUC1 originating from either bovine or human milk. Interspecies differences of the consensus repeat sequence in MUC1 and the physiological functions are discussed.
Pallesen
L T
LT
Protein Chemistry Laboratory, University of Aarhus, Denmark.
Andersen
M H
MH
Nielsen
R L
RL
Berglund
L
L
Petersen
T E
TE
Rasmussen
L K
LK
Rasmussen
J T
JT
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002845:Chromatography / Q000662:veterinary; D018076:DNA, Complementary / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D006017:Glycolipids / Q000032:analysis / Q000737:chemistry; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry; D006801:Humans; D066292:Lipid Droplets; D008892:Milk / Q000737:chemistry*; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry* / Q000302:isolation & purification; D015183:Restriction Mapping; D016415:Sequence Alignment; D020080:Tandem Repeat Sequences
D018076:DNA, Complementary; D006017:Glycolipids; D006023:Glycoproteins; D009077:Mucins; C110367:milk fat globule