Loop dynamics of thymidine diphosphate-rhamnose 3′-O-methyltransferase (CalS11), an enzyme in calicheamicin biosynthesis

• Published in: Structural dynamics (Melville, N.Y.) N.Y.), 2016-01, Vol.3 (1), p.012004-012004
• Main Authors: Han, Lu, Singh, Shanteri, Thorson, Jon S., Phillips, George N.
• Format: Article
• Language: English
• Subjects: Antibiotics; BASIC BIOLOGICAL SCIENCES; Biosynthesis; Carbohydrates; Catalysis; Crystal structure; Crystallography; Dynamic models; Dynamic structural analysis; Enediynes; Enzymes; Glycosylation; INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Molecular dynamics; Nucleotides; Protein structure; SPECIAL TOPIC: PROTEIN DYNAMICS; Substrates; Thymidine; X-ray diffraction
• ISSN: 2329-7778; 2329-7778
• DOI: 10.1063/1.4941368

Structure analysis and ensemble refinement of the apo-structure of thymidine diphosphate (TDP)-rhamnose 3′-O-methyltransferase reveal a gate for substrate entry and product release. TDP-rhamnose 3′-O-methyltransferase (CalS11) catalyses a 3′-O-methylation of TDP-rhamnose, an intermediate in the biosynthesis of enediyne antitumor antibiotic calicheamicin. CalS11 operates at the sugar nucleotide stage prior to glycosylation step. Here, we present the crystal structure of the apo form of CalS11 at 1.89 Å resolution. We propose that the L2 loop functions as a gate facilitating and/or providing specificity for substrate entry or promoting product release. Ensemble refinement analysis slightly improves the crystallographic refinement statistics and furthermore provides a compelling way to visualize the dynamic model of loop L2, supporting the understanding of its proposed role in catalysis.