Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite

Kadirvelraj, Renuka; Yang, Jeong-Yeh; Sanders, Justin H.; Liu, Lin; Ramiah, Annapoorani; Prabhakar, Pradeep Kumar; Boons, Geert-Jan; Wood, Zachary A.; Moremen, Kelley W.

doi:10.1073/pnas.1716988115

Human N-acetylglucosaminyltransferase II substrate recognition uses a modular architecture that includes a convergent exosite

Journal Article · Mon Apr 16 00:00:00 EDT 2018 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1716988115· OSTI ID:1464834

Kadirvelraj, Renuka ^[1]; Yang, Jeong-Yeh ^[1]; Sanders, Justin H. ^[1]; Liu, Lin ^[1]; Ramiah, Annapoorani ^[1]; Prabhakar, Pradeep Kumar ^[1]; Boons, Geert-Jan ^[1]; Wood, Zachary A. ^[1]; ^[1]

Univ. of Georgia, Athens, GA (United States)

Asn-linked oligosaccharides are extensively modified during transit through the secretory pathway, first by trimming of the nascent glycan chains and subsequently by initiating and extending multiple oligosaccharide branches from the trimannosyl glycan core. Trimming and branching pathway steps are highly ordered and hierarchal based on the precise substrate specificities of the individual biosynthetic enzymes. A key committed step in the synthesis of complex-type glycans is catalyzed by N-acetylglucosaminyltransferase II (MGAT2), an enzyme that generates the second GlcNAcβ1,2- branch from the trimannosyl glycan core using UDP-GlcNAc as the sugar donor. We determined the structure of human MGAT2 as a Mn²⁺-UDP donor analog complex and as a GlcNAcMan₃GlcNAc₂-Asn acceptor complex to reveal the structural basis for substrate recognition and catalysis. The enzyme exhibits a GT-A Rossmann-like fold that employs conserved divalent cation-dependent substrate interactions with the UDP-GlcNAc donor. MGAT2 interactions with the extended glycan acceptor are distinct from other related glycosyltransferases. These interactions are composed of a catalytic subsite that binds the Man-α1,6- monosaccharide acceptor and a distal exosite pocket that binds the GlcNAc-β1,2Man-α1,3Manβ- substrate “recognition arm.” Recognition arm interactions are similar to the enzyme–substrate interactions for Golgi α-mannosidase II, a glycoside hydrolase that acts just before MGAT2 in the Asn-linked glycan biosynthetic pathway. Here, these data suggest that substrate binding by MGAT2 employs both conserved and convergent catalytic subsite modules to provide substrate selectivity and catalysis. More broadly, the MGAT2 active-site architecture demonstrates how glycosyltransferases create complementary modular templates for regiospecific extension of glycan structures in mammalian cells.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: NIH

OSTI ID:: 1464834

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 18 Vol. 115; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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59 BASIC BIOLOGICAL SCIENCES
N-glycan processing
convergent evolution
exosite
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substrate recognition

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Cited By (6)

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