PMID- 10200178
OWN - NLM
STAT- MEDLINE
DCOM- 19990517
LR  - 20071114
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 15
DP  - 1999 Apr 13
TI  - Site-specific characterization of the N-linked glycans of murine prion protein by
      high-performance liquid chromatography/electrospray mass spectrometry and
      exoglycosidase digestions.
PG  - 4885-95
AB  - The murine prion protein PrP gene encodes a protein of 254 amino acids with two
      consensus sites for Asn-linked glycosylation at codons 180 and 196. A partial
      site-specific study of the N-linked glycans from hamster PrP has previously been 
      carried out by mass spectrometry [Stahl, N., Baldwin, M. A., Teplow, D. B., Hood,
      L., Gibson, B. W., Burlingame, A. L., and Prusiner, S. B. (1993) Biochemistry 32,
      1991-2002] and revealed that the glycosylation at Asn-181 (equivalent to mouse
      180) is heterogeneous, comprising over 30 glycoforms. The identification of the
      glycosylated peptide spanning Asn-197 was not reported. Recent technical advances
      in electrospray mass spectrometry now provide the sensitivity to detect low
      femtomole quantities of glycopeptides with >5000 mass resolution and 30 ppm mass 
      measurement [Medzihradszky, K. F., Besman, M. J., and Burlingame, A. L. (1998)
      Rapid Commun. Mass Spectrom. 12, 472-478]. This performance coupled with stepwise
      exoglycosidase digestion has been employed to establish the differential nature
      of the structural complexity (glycoforms) of the glycans at Asn-180 and Asn-196
      from a single strain infected with the ME7 strain. Some sixty structures have
      been found characterized by neutral and sialylated bi-, tri-, and tetraantennary 
      complex-type bearing outer-arm alpha(1-3)-fucosylation (the Lewisx and
      sialyl-Lewisx epitopes), core alpha(1,6) fucosylation, and the presence of
      terminal HexNAc residues. The Lewisx trisaccharide is the major nonreducing
      structure at Asn-180, and significant amounts of both Lewisx and sialyl Lewisx
      epitopes are observed at Asn-196. The abundance of the Lewisx and sialyl Lewisx
      epitopes on murine PrPSc may indicate a role for these structures in the normal
      function of PrPC or the pathophysiology of PrPSc.
FAU - Stimson, E
AU  - Stimson E
AD  - Ludwig Institute for Cancer Research, 91 Riding House Street, London, U.K.
FAU - Hope, J
AU  - Hope J
FAU - Chong, A
AU  - Chong A
FAU - Burlingame, A L
AU  - Burlingame AL
LA  - eng
GR  - RR 01614/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Polysaccharides)
RN  - 0 (Prions)
RN  - 7006-34-0 (Asparagine)
RN  - EC 3.2.1.- (Glycoside Hydrolases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Asparagine/metabolism
MH  - Carbohydrate Conformation
MH  - Carbohydrate Sequence
MH  - Chromatography, High Pressure Liquid
MH  - Glycoside Hydrolases/*metabolism
MH  - Hydrolysis
MH  - Mass Spectrometry
MH  - Mice
MH  - Molecular Sequence Data
MH  - Polysaccharides/chemistry/*metabolism
MH  - Prions/chemistry/*metabolism
EDAT- 1999/04/14 00:00
MHDA- 1999/04/14 00:01
CRDT- 1999/04/14 00:00
PHST- 1999/04/14 00:00 [pubmed]
PHST- 1999/04/14 00:01 [medline]
PHST- 1999/04/14 00:00 [entrez]
AID - 10.1021/bi982330q [doi]
AID - bi982330q [pii]
PST - ppublish
SO  - Biochemistry. 1999 Apr 13;38(15):4885-95. doi: 10.1021/bi982330q.