2JF6

Structure of inactive mutant of Strictosidine Glucosidase in complex with strictosidine

Summary for 2JF6

• Entry DOI: 10.2210/pdb2jf6/pdb
• Related: 2JF7
• Descriptor: STRICTOSIDINE-O-BETA-D-GLUCOSIDASE, METHYL (2S,3R,4S)-3-ETHYL-2-(BETA-D-GLUCOPYRANOSYLOXY)-4-[(1S)-2,3,4,9-TETRAHYDRO-1H-BETA-CARBOLIN-1-YLMETHYL]-3,4-DIHYDRO-2H-PYRAN-5-CARBOXYLATE (3 entities in total)
• Functional Keywords: alkaloid, hydrolase
• Biological source: RAUVOLFIA SERPENTINA (SERPENTWOOD)
• Total number of polymer chains: 2
• Total formula weight: 122983.96

Authors

Barleben, L.,Panjikar, S.,Ruppert, M.,Koepke, J.,Stockigt, J. (deposition date: 2007-01-26, release date: 2008-02-05, Last modification date: 2023-12-13)

Primary citation

Barleben, L.,Panjikar, S.,Ruppert, M.,Koepke, J.,Stockigt, J.
Molecular Architecture of Strictosidine Glucosidase - the Gateway to the Biosynthesis of the Monoterpenoid Indole Alkaloid Family
Plant Cell, 19:2886-, 2007

PubMed Abstract: Strictosidine beta-D-glucosidase (SG) follows strictosidine synthase (STR1) in the production of the reactive intermediate required for the formation of the large family of monoterpenoid indole alkaloids in plants. This family is composed of approximately 2000 structurally diverse compounds. SG plays an important role in the plant cell by activating the glucoside strictosidine and allowing it to enter the multiple indole alkaloid pathways. Here, we report detailed three-dimensional information describing both native SG and the complex of its inactive mutant Glu207Gln with the substrate strictosidine, thus providing a structural characterization of substrate binding and identifying the amino acids that occupy the active site surface of the enzyme. Structural analysis and site-directed mutagenesis experiments demonstrate the essential role of Glu-207, Glu-416, His-161, and Trp-388 in catalysis. Comparison of the catalytic pocket of SG with that of other plant glucosidases demonstrates the structural importance of Trp-388. Compared with all other glucosidases of plant, bacterial, and archaeal origin, SG's residue Trp-388 is present in a unique structural conformation that is specific to the SG enzyme. In addition to STR1 and vinorine synthase, SG represents the third structural example of enzymes participating in the biosynthetic pathway of the Rauvolfia alkaloid ajmaline. The data presented here will contribute to deciphering the structure and reaction mechanism of other higher plant glucosidases.

PubMed: 17890378
DOI: 10.1105/TPC.106.045682

Experimental method

X-RAY DIFFRACTION (2.82 Å)