Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity

Johannes W Pedersen; Eric P Bennett; Katrine T-B G Schjoldager; Morten Meldal; Andreas P Holmér; Ola Blixt; Emiliano Cló; Steven B Levery; Henrik Clausen; Hans H Wandall

doi:10.1074/jbc.M111.273722

Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity

Johannes W Pedersen, Eric P Bennett, Katrine T-B G Schjoldager, Morten Meldal, Andreas P Holmér, Ola Blixt, Emiliano Cló, Steven B Levery, Henrik Clausen, Hans H Wandall

41 Citationer (Scopus)

Abstract

UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferases (GalNAc-Ts) constitute a family of up to 20 transferases that initiate mucin-type O-glycosylation. The transferases are structurally composed of catalytic and lectin domains. Two modes have been identified for the selection of glycosylation sites by GalNAc-Ts: confined sequence recognition by the catalytic domain alone, and concerted recognition of acceptor sites and adjacent GalNAc-glycosylated sites by the catalytic and lectin domains, respectively. Thus far, only the catalytic domain has been shown to have peptide sequence specificity, whereas the primary function of the lectin domain is to increase affinity to previously glycosylated substrates. Whether the lectin domain also has peptide sequence selectivity has remained unclear. Using a glycopeptide array with a library of synthetic and recombinant glycopeptides based on sequences of mucins MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC7 as well as a random glycopeptide bead library, we examined the binding properties of four different lectin domains. The lectin domains of GalNAc-T1, -T2, -T3, and -T4 bound different subsets of small glycopeptides. These results indicate an additional level of complexity in the initiation step of O-glycosylation by GalNAc-Ts.

Originalsprog	Engelsk
Tidsskrift	The Journal of Biological Chemistry
Vol/bind	286
Udgave nummer	37
Sider (fra-til)	32684-96
Antal sider	13
ISSN	1083-351X
DOI	https://doi.org/10.1074/jbc.M111.273722
Status	Udgivet - 16 sep. 2011

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10.1074/jbc.M111.273722

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title = "Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity",

abstract = "UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferases (GalNAc-Ts) constitute a family of up to 20 transferases that initiate mucin-type O-glycosylation. The transferases are structurally composed of catalytic and lectin domains. Two modes have been identified for the selection of glycosylation sites by GalNAc-Ts: confined sequence recognition by the catalytic domain alone, and concerted recognition of acceptor sites and adjacent GalNAc-glycosylated sites by the catalytic and lectin domains, respectively. Thus far, only the catalytic domain has been shown to have peptide sequence specificity, whereas the primary function of the lectin domain is to increase affinity to previously glycosylated substrates. Whether the lectin domain also has peptide sequence selectivity has remained unclear. Using a glycopeptide array with a library of synthetic and recombinant glycopeptides based on sequences of mucins MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC7 as well as a random glycopeptide bead library, we examined the binding properties of four different lectin domains. The lectin domains of GalNAc-T1, -T2, -T3, and -T4 bound different subsets of small glycopeptides. These results indicate an additional level of complexity in the initiation step of O-glycosylation by GalNAc-Ts.",

author = "Pedersen, {Johannes W} and Bennett, {Eric P} and Schjoldager, {Katrine T-B G} and Morten Meldal and Holm{\'e}r, {Andreas P} and Ola Blixt and Emiliano Cl{\'o} and Levery, {Steven B} and Henrik Clausen and Wandall, {Hans H}",

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T1 - Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity

AU - Pedersen, Johannes W

AU - Bennett, Eric P

AU - Schjoldager, Katrine T-B G

AU - Meldal, Morten

AU - Holmér, Andreas P

AU - Blixt, Ola

AU - Cló, Emiliano

AU - Levery, Steven B

AU - Clausen, Henrik

AU - Wandall, Hans H

PY - 2011/9/16

Y1 - 2011/9/16

N2 - UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferases (GalNAc-Ts) constitute a family of up to 20 transferases that initiate mucin-type O-glycosylation. The transferases are structurally composed of catalytic and lectin domains. Two modes have been identified for the selection of glycosylation sites by GalNAc-Ts: confined sequence recognition by the catalytic domain alone, and concerted recognition of acceptor sites and adjacent GalNAc-glycosylated sites by the catalytic and lectin domains, respectively. Thus far, only the catalytic domain has been shown to have peptide sequence specificity, whereas the primary function of the lectin domain is to increase affinity to previously glycosylated substrates. Whether the lectin domain also has peptide sequence selectivity has remained unclear. Using a glycopeptide array with a library of synthetic and recombinant glycopeptides based on sequences of mucins MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC7 as well as a random glycopeptide bead library, we examined the binding properties of four different lectin domains. The lectin domains of GalNAc-T1, -T2, -T3, and -T4 bound different subsets of small glycopeptides. These results indicate an additional level of complexity in the initiation step of O-glycosylation by GalNAc-Ts.

AB - UDP-GalNAc:polypeptide α-N-acetylgalactosaminyltransferases (GalNAc-Ts) constitute a family of up to 20 transferases that initiate mucin-type O-glycosylation. The transferases are structurally composed of catalytic and lectin domains. Two modes have been identified for the selection of glycosylation sites by GalNAc-Ts: confined sequence recognition by the catalytic domain alone, and concerted recognition of acceptor sites and adjacent GalNAc-glycosylated sites by the catalytic and lectin domains, respectively. Thus far, only the catalytic domain has been shown to have peptide sequence specificity, whereas the primary function of the lectin domain is to increase affinity to previously glycosylated substrates. Whether the lectin domain also has peptide sequence selectivity has remained unclear. Using a glycopeptide array with a library of synthetic and recombinant glycopeptides based on sequences of mucins MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC7 as well as a random glycopeptide bead library, we examined the binding properties of four different lectin domains. The lectin domains of GalNAc-T1, -T2, -T3, and -T4 bound different subsets of small glycopeptides. These results indicate an additional level of complexity in the initiation step of O-glycosylation by GalNAc-Ts.

U2 - 10.1074/jbc.M111.273722

DO - 10.1074/jbc.M111.273722

M3 - Journal article

C2 - 21768105

SN - 0021-9258

VL - 286

SP - 32684

EP - 32696

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 37

ER -

Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity

Abstract

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