4AP6

Crystal structure of human POFUT2 E54A mutant in complex with GDP- fucose

Summary for 4AP6

• Entry DOI: 10.2210/pdb4ap6/pdb
• Related: 4AP5
• Descriptor: GDP-FUCOSE PROTEIN O-FUCOSYLTRANSFERASE 2, GUANOSINE-5'-DIPHOSPHATE-BETA-L-FUCOPYRANOSE, 2-acetamido-2-deoxy-beta-D-glucopyranose, ... (5 entities in total)
• Functional Keywords: transferase, gt-b, gt68
• Biological source: HOMO SAPIENS (HUMAN)
• Cellular location: Endoplasmic reticulum (By similarity): Q9Y2G5
• Total number of polymer chains: 4
• Total formula weight: 202042.27

Authors

Chen, C.,Keusch, J.J.,Klein, D.,Hess, D.,Hofsteenge, J.,Gut, H. (deposition date: 2012-03-30, release date: 2012-08-08, Last modification date: 2024-11-13)

Primary citation

Chen, C.I.,Keusch, J.J.,Klein, D.,Hess, D.,Hofsteenge, J.,Gut, H.
Structure of Human Pofut2: Insights Into Thrombospondin Type 1 Repeat Fold and O-Fucosylation.
Embo J., 31:3183-, 2012

PubMed Abstract: Protein O-fucosylation is a post-translational modification found on serine/threonine residues of thrombospondin type 1 repeats (TSR). The fucose transfer is catalysed by the protein O-fucosyltransferase 2 (POFUT2) and >40 human proteins contain the TSR consensus sequence for POFUT2-dependent fucosylation. To better understand O-fucosylation on TSR, we carried out a structural and functional analysis of human POFUT2 and its TSR substrate. Crystal structures of POFUT2 reveal a variation of the classical GT-B fold and identify sugar donor and TSR acceptor binding sites. Structural findings are correlated with steady-state kinetic measurements of wild-type and mutant POFUT2 and TSR and give insight into the catalytic mechanism and substrate specificity. By using an artificial mini-TSR substrate, we show that specificity is not primarily encoded in the TSR protein sequence but rather in the unusual 3D structure of a small part of the TSR. Our findings uncover that recognition of distinct conserved 3D fold motifs can be used as a mechanism to achieve substrate specificity by enzymes modifying completely folded proteins of very wide sequence diversity and biological function.

PubMed: 22588082
DOI: 10.1038/EMBOJ.2012.143

Experimental method

X-RAY DIFFRACTION (3.401 Å)