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Crystallization and X-ray Diffraction Analysis of a Putative Bacterial Class I Labdane-Related Diterpene Synthase

Crystallization and X-ray Diffraction Analysis of a Putative Bacterial Class I Labdane-Related Diterpene Synthase

Hugo Serrano-Posada, Sara Centeno-Leija, Sonia Rojas-Trejo, Vivian Stojanoff, Romina Rodríguez-Sanoja, Enrique Rudiño-Piñera, Sergio Sánchez

Acta Crystallogr F Struct Biol Commun. 2015 Sep;71(Pt 9):1194-9.

PMID: 26323307

Abstract:

Labdane-related diterpenoids are natural products with potential pharmaceutical applications that are rarely found in bacteria. Here, a putative class I labdane-related diterpene synthase (LrdC) identified by genome mining in a streptomycete was successfully crystallized using the microbatch method. Crystals of the LrdC enzyme were obtained in a holo form with its natural cofactor Mg(2+) (LrdC-Mg(2+)) and in complex with inorganic pyrophosphate (PPi) (LrdC-Mg(2+)-PPi). Crystals of native LrdC-Mg(2+) diffracted to 2.50 Å resolution and belonged to the trigonal space group P3221, with unit-cell parameters a = b = 107.1, c = 89.2 Å. Crystals of the LrdC-Mg(2+)-PPi complex grown in the same conditions as the native enzyme with PEG 8000 diffracted to 2.36 Å resolution and also belonged to the trigonal space group P3221. Crystals of the LrdC-Mg(2+)-PPi complex grown in a second crystallization condition with PEG 3350 diffracted to 2.57 Å resolution and belonged to the monoclinic space group P21, with unit-cell parameters a = 49.9, b = 104.1, c = 66.5 Å, β = 111.4°. The structure was determined by the single-wavelength anomalous dispersion (SAD) technique using the osmium signal from a potassium hexachloroosmate (IV) derivative.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP16871606	Potassium hexachloroosmate(IV)		16871-60-6	Price

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