Structural rearrangements of sucrose phosphorylase from Bifidobacterium adolescentis during sucrose conversion
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Structural rearrangements of sucrose phosphorylase from Bifidobacterium adolescentis during sucrose conversion. / Mirza, Osman; Henriksen, Lars Skov; Sprogøe, Desiree; van den Broek, Lambertus A M; Beldman, Gerrit; Kastrup, Jette S; Gajhede, Michael.
In: Journal of Biological Chemistry, Vol. 281, No. 46, 2006, p. 35576-84.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Structural rearrangements of sucrose phosphorylase from Bifidobacterium adolescentis during sucrose conversion
AU - Mirza, Osman
AU - Henriksen, Lars Skov
AU - Sprogøe, Desiree
AU - van den Broek, Lambertus A M
AU - Beldman, Gerrit
AU - Kastrup, Jette S
AU - Gajhede, Michael
PY - 2006
Y1 - 2006
N2 - The reaction mechanism of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) was studied by site-directed mutagenesis and x-ray crystallography. An inactive mutant of BiSP (E232Q) was co-crystallized with sucrose. The structure revealed a substrate-binding mode comparable with that seen in other related sucrose-acting enzymes. Wild-type BiSP was also crystallized in the presence of sucrose. In the dimeric structure, a covalent glucosyl intermediate was formed in one molecule of the BiSP dimer, and after hydrolysis of the glucosyl intermediate, a beta-D-glucose product complex was formed in the other molecule. Although the overall structure of the BiSP-glucosyl intermediate complex is similar to that of the BiSP(E232Q)-sucrose complex, the glucose complex discloses major differences in loop conformations. Two loops (residues 336-344 and 132-137) in the proximity of the active site move up to 16 and 4 A, respectively. On the basis of these findings, we have suggested a reaction cycle that takes into account the large movements in the active-site entrance loops.
AB - The reaction mechanism of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) was studied by site-directed mutagenesis and x-ray crystallography. An inactive mutant of BiSP (E232Q) was co-crystallized with sucrose. The structure revealed a substrate-binding mode comparable with that seen in other related sucrose-acting enzymes. Wild-type BiSP was also crystallized in the presence of sucrose. In the dimeric structure, a covalent glucosyl intermediate was formed in one molecule of the BiSP dimer, and after hydrolysis of the glucosyl intermediate, a beta-D-glucose product complex was formed in the other molecule. Although the overall structure of the BiSP-glucosyl intermediate complex is similar to that of the BiSP(E232Q)-sucrose complex, the glucose complex discloses major differences in loop conformations. Two loops (residues 336-344 and 132-137) in the proximity of the active site move up to 16 and 4 A, respectively. On the basis of these findings, we have suggested a reaction cycle that takes into account the large movements in the active-site entrance loops.
KW - Bifidobacterium
KW - Binding Sites
KW - Carbohydrate Conformation
KW - Glucosyltransferases
KW - Models, Molecular
KW - Protein Conformation
KW - Sucrose
U2 - 10.1074/jbc.M605611200
DO - 10.1074/jbc.M605611200
M3 - Journal article
C2 - 16990265
VL - 281
SP - 35576
EP - 35584
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 46
ER -
ID: 40766583