Toward a blueprint for UDP-glucose pyrophosphorylase structure/function properties: homology-modeling analyses.
UDP-glucose pyrophosphorylase (UGPase) is an important enzyme of synthesis of sucrose, cellulose, and several other polysaccharides in all plants. The protein is evolutionarily conserved among eukaryotes, but has little relation, aside from its catalytic reaction, to UGPases of prokaryotic origin. Using protein homology modeling strategy, 3D structures for barley, poplar, and Arabidopsis UGPases have been derived, based on recently published crystal structure of human UDP-N-acetylglucosamine pyrophosphorylase. The derived 3D structures correspond to a bowl-shaped protein with the active site at a central groove, and a C-terminal domain that includes a loop (I-loop) possibly involved in dimerization. Data on a plethora of earlier described UGPase mutants from a variety of eukaryotic organisms have been revisited, and we have, in most cases, verified the role of each mutation in enzyme catalysis/regulation/structural integrity. We have also found that one of two alternatively spliced forms of poplar UGPase has a very short I-loop, suggesting differences in oligomerization ability of the two isozymes. The derivation of the structural model for plant UGPase should serve as a useful blueprint for further function/structure studies on this protein.
Geisler
Matt
M
Department of Plant Physiology, Umeå Plant Science Centre, Umeå University, 901 87 Umeå, Sweden.
Wilczynska
Malgorzata
M
Karpinski
Stanislaw
S
Kleczkowski
Leszek A
LA
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D005057:Eukaryotic Cells / Q000201:enzymology; D014644:Genetic Variation; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D010944:Plants / Q000201:enzymology* / Q000235:genetics; D032107:Populus / Q000201:enzymology / Q000235:genetics; D011387:Prokaryotic Cells / Q000201:enzymology; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000737:chemistry / Q000235:genetics*
D007527:Isoenzymes; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase