3VGF

Crystal structure of glycosyltrehalose trehalohydrolase (D252S) complexed with maltotriosyltrehalose

Summary for 3VGF

• Entry DOI: 10.2210/pdb3vgf/pdb
• Related: 1EH9 3VGC 3VGD 3VGE 3VGG 3VGH
• Related PRD ID: PRD_900009
• Descriptor: Malto-oligosyltrehalose trehalohydrolase, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose, CITRATE ANION, ... (5 entities in total)
• Functional Keywords: alpha/beta barrel, trehalose, trehalohydrolase, alpha-amylase, hydrolase
• Biological source: Sulfolobus solfataricus
• Cellular location: Cytoplasm: Q55088
• Total number of polymer chains: 1
• Total formula weight: 65959.82

Authors

Okazaki, N.,Tamada, T.,Feese, M.D.,Kato, M.,Miura, Y.,Komeda, T.,Kobayashi, K.,Kondo, K.,Kuroki, R. (deposition date: 2011-08-09, release date: 2012-06-20, Last modification date: 2023-11-08)

Primary citation

Okazaki, N.,Tamada, T.,Feese, M.D.,Kato, M.,Miura, Y.,Komeda, T.,Kobayashi, K.,Kondo, K.,Blaber, M.,Kuroki, R.
Substrate recognition mechanism of a glycosyltrehalose trehalohydrolase from Sulfolobus solfataricus KM1.
Protein Sci., 21:539-552, 2012

PubMed Abstract: Glycosyltrehalose trehalohydrolase (GTHase) is an α-amylase that cleaves the α-1,4 bond adjacent to the α-1,1 bond of maltooligosyltrehalose to release trehalose. To investigate the catalytic and substrate recognition mechanisms of GTHase, two residues, Asp252 (nucleophile) and Glu283 (general acid/base), located at the catalytic site of GTHase were mutated (Asp252→Ser (D252S), Glu (D252E) and Glu283→Gln (E283Q)), and the activity and structure of the enzyme were investigated. The E283Q, D252E, and D252S mutants showed only 0.04, 0.03, and 0.6% of enzymatic activity against the wild-type, respectively. The crystal structure of the E283Q mutant GTHase in complex with the substrate, maltotriosyltrehalose (G3-Tre), was determined to 2.6-Å resolution. The structure with G3-Tre indicated that GTHase has at least five substrate binding subsites and that Glu283 is the catalytic acid, and Asp252 is the nucleophile that attacks the C1 carbon in the glycosidic linkage of G3-Tre. The complex structure also revealed a scheme for substrate recognition by GTHase. Substrate recognition involves two unique interactions: stacking of Tyr325 with the terminal glucose ring of the trehalose moiety and perpendicularly placement of Trp215 to the pyranose rings at the subsites -1 and +1 glucose.

PubMed: 22334583
DOI: 10.1002/pro.2039

Experimental method

X-RAY DIFFRACTION (2.3 Å)