The beginnings of mucin biosynthesis:: The crystal structure of UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferase-T1

被引:120
作者
Fritz, TA
Hurley, JH
Trinh, LB
Shiloach, J
Tabak, LA [1 ]
机构
[1] NIDDKD, Sect Biol Chem, NIH, Dept Hlth & Human Serv, Bethesda, MD 20892 USA
[2] NIDDKD, Mol Biol Lab, NIH, Dept Hlth & Human Serv, Bethesda, MD 20892 USA
[3] NIDDKD, Biotechnol Unit, NIH, Dept Hlth & Human Serv, Bethesda, MD 20892 USA
关键词
glycosyltransferase; mucin;
D O I
10.1073/pnas.0405657101
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
UDP-GaINAc:polypeptide alpha-N-acetylgalactosaminyltransferases (ppGaNTases) initiate the formation of mucin-type, O-linked glycans by catalyzing the transfer of alpha-N-acetylgalactosamine from UDP-GaINAc to Ser or Thr residues of core proteins to form the Tin antigen (GalNAc-alpha-1-O-Ser/Thr). ppGaNTases are unique among glycosyltransferases in containing a C-terminal lectin domain. We present the x-ray crystal structure of a ppGaNTase, murine ppGaNTase-T1, and show that it folds to form distinct catalytic and lectin domains. The association of the two domains forms a large cleft in the surface of the enzyme that contains a Mn2+ ion complexed by invariant D209 and H211 of the "DXH" motif and by invariant H344. Each of the three potential lectin domain carbohydrate-binding sites (alpha, beta, and gamma) is located on the active-site face of the enzyme, suggesting a mechanism by which the transferase may accommodate multiple conformations of glycosylated acceptor substrates. A model of a mucin 1 glycopepticle substrate bound to the enzyme shows that the spatial separation between the lectin alpha site and a modeled active site UDP-GaINAc is consistent with the in vitro pattern of glycosylation observed for this peptide catalyzed by ppGaNTase-T1. The structure also provides a template for the larger ppGaNTase family, and homology models of several ppGaNTase isoforms predict dramatically different surface chemistries consistent with isoform-selective acceptor substrate recognition.
引用
收藏
页码:15307 / 15312
页数:6
相关论文
共 51 条
[41]   Automated MAD and MIR structure solution [J].
Terwilliger, TC ;
Berendzen, J .
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 1999, 55 :849-861
[42]   Automated main-chain model building by template matching and iterative fragment extension [J].
Terwilliger, TC .
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2003, 59 :38-44
[43]   Maximum-likelihood density modification [J].
Terwilliger, TC .
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2000, 56 :965-972
[44]   Glycopeptide N-acetylgalactosaminyltransferase specificities for O-glycosylated sites on MUC5AC mucin motif peptides [J].
Tetaert, D ;
Ten Hagen, KG ;
Richet, C ;
Boersma, A ;
Gagnon, J ;
Degand, P .
BIOCHEMICAL JOURNAL, 2001, 357 (01) :313-320
[45]   Mutations in GALNT3, encoding a protein involved in O-linked glycosylation, cause familial tumoral calcinosis [J].
Topaz, O ;
Shurman, DI ;
Bergman, R ;
Indelman, M ;
Ratajczak, P ;
Mizrachi, M ;
Khamaysi, Z ;
Behar, D ;
Petronius, D ;
Friedman, V ;
Zelikovic, I ;
Raimer, S ;
Metzker, A ;
Richard, G ;
Sprecher, E .
NATURE GENETICS, 2004, 36 (06) :579-581
[46]   X-ray crystal structure of rabbit N-acetylglucosaminyltransferase I:: catalytic mechanism and a new protein superfamily [J].
Ünligil, UM ;
Zhou, SH ;
Yuwaraj, S ;
Sarkar, M ;
Schachter, H ;
Rini, JM .
EMBO JOURNAL, 2000, 19 (20) :5269-5280
[47]   Substrate specificities of three members of the human UDP-N-acetyl-alpha-D-galactosamine:Polypeptide N-acetylgalactosaminyltransferase family, GalNAc-T1, -T2, and -T3 [J].
Wandall, HH ;
Hassan, H ;
Mirgorodskaya, E ;
Kristensen, AK ;
Roepstorff, P ;
Bennett, EP ;
Nielsen, PA ;
Hollingsworth, MA ;
Burchell, J ;
TaylorPapadimitriou, J ;
Clausen, H .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (38) :23503-23514
[48]   Identification of essential histidine residues in UDP-N-acetyl-D-galactosamine: polypeptide N-acetylgalactosaminyltransferase-T1 [J].
Wragg, S ;
Hagen, FK ;
Tabak, LA .
BIOCHEMICAL JOURNAL, 1997, 328 :193-197
[49]   Defective angiogenesis and fatal embryonic hemorrhage in mice lacking core 1-derived O-glycans [J].
Xia, LJ ;
Ju, TZ ;
Westmuckett, A ;
An, GY ;
Ivanciu, L ;
McDaniel, JM ;
Lupu, F ;
Cummings, RD ;
McEver, RP .
JOURNAL OF CELL BIOLOGY, 2004, 164 (03) :451-459
[50]   Organization of golgi glycosyltransferases in membranes: Complexity via complexes [J].
Young, WW .
JOURNAL OF MEMBRANE BIOLOGY, 2004, 198 (01) :1-13