PMID- 10373415
OWN - NLM
STAT- MEDLINE
DCOM- 19990715
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 26
DP  - 1999 Jun 25
TI  - High density O-glycosylation on tandem repeat peptide from secretory MUC1 of T47D
      breast cancer cells.
PG  - 18165-72
AB  - The site-specific O-glycosylation of MUC1 tandem repeat peptides from secretory
      mucin of T47D breast cancer cells was analyzed. After affinity isolation on
      immobilized BC3 antibody, MUC1 was partially deglycosylated by enzymatic
      treatment with alpha-sialidase/beta-galactosidase and fragmented by proteolytic
      cleavage with the Arg-C-specific endopeptidase clostripain. The PAP20
      glycopeptides were isolated by reversed phase high pressure liquid chromatography
      and subjected to the structural analyses by quadrupole time-of-flight
      electrospray ionization mass spectrometry and to the sequencing by Edman
      degradation. All five positions of the repeat peptide were revealed as
      O-glycosylation targets in the tumor cell, including the Thr within the DTR
      motif. The degree of substitution was estimated to average 4.8 glycans per
      repeat, which compares to 2.6 glycosylated sites per repeat for the mucin from
      milk (Muller, S., Goletz, S., Packer, N., Gooley, A. A., Lawson, A. M., and
      Hanisch, F.-G. (1997) J. Biol. Chem. 272, 24780-24793). In addition to a
      modification by glycosylation, the immunodominant DTR motif on T47D-MUC1 is
      altered by amino acid replacements (PAPGSTAPAAHGVTSAPESR), which were revealed in
      about 50% of PAP20 peptides. The high incidence of these replacements and their
      detection also in other cancer cell lines imply that the conserved tandem repeat 
      domain of MUC1 is polymorphic with respect to the peptide sequence.
FAU - Muller, S
AU  - Muller S
AD  - Institute of Biochemistry, Medical Faculty of the University,
      Joseph-Stelzmann-Strasse 52, 50931 Koln, Germany.
FAU - Alving, K
AU  - Alving K
FAU - Peter-Katalinic, J
AU  - Peter-Katalinic J
FAU - Zachara, N
AU  - Zachara N
FAU - Gooley, A A
AU  - Gooley AA
FAU - Hanisch, F G
AU  - Hanisch FG
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Mucin-1)
RN  - EC 3.4.- (Carboxypeptidases)
RN  - EC 3.4.16.5 (CTSA protein, human)
RN  - EC 3.4.16.5 (Cathepsin A)
RN  - EC 3.4.22.- (Cysteine Endopeptidases)
RN  - EC 3.4.22.8 (clostripain)
SB  - IM
MH  - Amino Acid Sequence
MH  - Blotting, Western
MH  - Breast Neoplasms/*chemistry
MH  - Carbohydrate Sequence
MH  - Carboxypeptidases/metabolism
MH  - Cathepsin A
MH  - Chromatography, High Pressure Liquid
MH  - Cysteine Endopeptidases/metabolism
MH  - Female
MH  - Glycosylation
MH  - Humans
MH  - Mass Spectrometry
MH  - Molecular Sequence Data
MH  - Mucin-1/*chemistry
MH  - Tumor Cells, Cultured
EDAT- 1999/06/22 00:00
MHDA- 1999/06/22 00:01
CRDT- 1999/06/22 00:00
PHST- 1999/06/22 00:00 [pubmed]
PHST- 1999/06/22 00:01 [medline]
PHST- 1999/06/22 00:00 [entrez]
AID - 10.1074/jbc.274.26.18165 [doi]
AID - S0021-9258(19)87141-7 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Jun 25;274(26):18165-72. doi: 10.1074/jbc.274.26.18165.