PMID- 32518939
OWN - NLM
STAT- MEDLINE
DCOM- 20220117
LR  - 20220117
IS  - 1460-2423 (Electronic)
IS  - 0959-6658 (Linking)
VI  - 31
IP  - 1
DP  - 2021 Jan 9
TI  - The single EGF-like domain of mouse PAMR1 is modified by O-Glucose, O-Fucose and 
      O-GlcNAc.
PG  - 55-68
LID - 10.1093/glycob/cwaa051 [doi]
AB  - Epidermal growth factor-like domains (EGF-LDs) of membrane and secreted proteins 
      can be modified by N-glycans and/or potentially elongated O-linked 
      monosaccharides such as O-glucose (O-Glc) found at two positions (O-Glc 1 and 
      O-Glc2), O-fucose (O-Fuc) and O-N-acetylglucosamine (O-GlcNAc). The presence of 
      three O-linked sugars within the same EGF-LD, such as in EGF-LD 20 of NOTCH1, has 
      rarely been evidenced. We searched in KEGG GENES database to list mouse and human 
      proteins with an EGF-LD sequence including one, two, three or four potential 
      O-glycosylation consensus sites. Among the 129 murine retrieved proteins, most 
      had predicted O-fucosylation and/or O-GlcNAcylation sites. Around 68% of EGF-LDs 
      were subjected to only one O-linked sugar modification and near 5% to three 
      modifications. Among these latter, we focused on the peptidase domain-containing 
      protein associated with muscle regeneration 1 (PAMR1), having only one EGF-LD. To 
      test the ability of this domain to be glycosylated, a correctly folded EGF-LD was 
      produced in Escherichia coli periplasm, purified and subjected to in vitro 
      incubations with the recombinant O-glycosyltransferases POGLUT1, POFUT1 and EOGT, 
      adding O-Glc1, O-Fuc and O-GlcNAc, respectively. Using click chemistry and mass 
      spectrometry, isolated PAMR1 EGF-LD was demonstrated to be modified by the three 
      O-linked sugars. Their presence was individually confirmed on EGF-LD of 
      full-length mouse recombinant PAMR1, with at least some molecules modified by 
      both O-Glc1 and O-Fuc. Overall, these results are consistent with the presence of 
      a triple O-glycosylated EGF-LD in mouse PAMR1.
CI  - (c) The Author(s) 2020. Published by Oxford University Press. All rights reserved. 
      For permissions, please e-mail: journals.permissions@oup.com.
FAU - Pennarubia, Florian
AU  - Pennarubia F
AD  - University of Limoges, PEIRENE, EA 7500, Glycosylation and Cell Differentiation, 
      F-87060 Limoges, France.
FAU - Germot, Agnes
AU  - Germot A
AD  - University of Limoges, PEIRENE, EA 7500, Glycosylation and Cell Differentiation, 
      F-87060 Limoges, France.
FAU - Pinault, Emilie
AU  - Pinault E
AD  - University of Limoges, PEIRENE, EA 7500, Glycosylation and Cell Differentiation, 
      F-87060 Limoges, France.
AD  - University of Limoges, BISCEm, US 42 INSERM - UMS 2015 CNRS, Mass Spectrometry 
      Platform, F-87025 Limoges, France.
FAU - Maftah, Abderrahman
AU  - Maftah A
AD  - University of Limoges, PEIRENE, EA 7500, Glycosylation and Cell Differentiation, 
      F-87060 Limoges, France.
FAU - Legardinier, Sebastien
AU  - Legardinier S
AD  - University of Limoges, PEIRENE, EA 7500, Glycosylation and Cell Differentiation, 
      F-87060 Limoges, France.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Glycobiology
JT  - Glycobiology
JID - 9104124
RN  - 0 (Recombinant Proteins)
RN  - 28RYY2IV3F (Fucose)
RN  - 62229-50-9 (Epidermal Growth Factor)
RN  - EC 2.4.1.- (N-Acetylglucosaminyltransferases)
RN  - EC 3.4.- (Serine Proteases)
RN  - EC 3.4.21.- (Pamr1 protein, mouse)
RN  - IY9XDZ35W2 (Glucose)
RN  - V956696549 (Acetylglucosamine)
SB  - IM
MH  - Acetylglucosamine/chemistry/*metabolism
MH  - Animals
MH  - Databases, Protein
MH  - Epidermal Growth Factor/chemistry/isolation & purification/*metabolism
MH  - Fucose/chemistry/*metabolism
MH  - Glucose/chemistry/*metabolism
MH  - Humans
MH  - Mice
MH  - N-Acetylglucosaminyltransferases/chemistry/*metabolism
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Serine Proteases/chemistry/*metabolism
OTO - NOTNLM
OT  - EGF-LD
OT  - EOGT
OT  - PAMR1
OT  - POFUT1
OT  - POGLUT
EDAT- 2020/06/11 06:00
MHDA- 2022/01/18 06:00
CRDT- 2020/06/11 06:00
PHST- 2020/04/19 00:00 [received]
PHST- 2020/05/22 00:00 [revised]
PHST- 2020/05/28 00:00 [accepted]
PHST- 2020/06/11 06:00 [pubmed]
PHST- 2022/01/18 06:00 [medline]
PHST- 2020/06/11 06:00 [entrez]
AID - 5849841 [pii]
AID - 10.1093/glycob/cwaa051 [doi]
PST - ppublish
SO  - Glycobiology. 2021 Jan 9;31(1):55-68. doi: 10.1093/glycob/cwaa051.