3SBM

Trans-acting transferase from Disorazole synthase in complex with Acetate

Summary for 3SBM

• Entry DOI: 10.2210/pdb3sbm/pdb
• Descriptor: DisD protein, ACETATE ION, HEXAETHYLENE GLYCOL, ... (4 entities in total)
• Functional Keywords: transferase
• Biological source: Sorangium cellulosum
• Total number of polymer chains: 1
• Total formula weight: 30994.38

Authors

Khosla, C.,Mathews, I.I.,Wong, F.T.,Jin, X. (deposition date: 2011-06-06, release date: 2011-07-13, Last modification date: 2023-09-13)

Primary citation

Wong, F.T.,Jin, X.,Mathews, I.I.,Cane, D.E.,Khosla, C.
Structure and Mechanism of the trans-Acting Acyltransferase from the Disorazole Synthase.
Biochemistry, 50:6539-6548, 2011

PubMed Abstract: The 1.51 Å resolution X-ray crystal structure of the trans-acyltransferase (AT) from the "AT-less" disorazole synthase (DSZS) and that of its acetate complex at 1.35 Å resolution are reported. Separately, comprehensive alanine-scanning mutagenesis of one of its acyl carrier protein substrates (ACP1 from DSZS) led to the identification of a conserved Asp45 residue on the ACP, which contributes to the substrate specificity of this unusual enzyme. Together, these experimental findings were used to derive a model for the selective association of the DSZS AT and its ACP substrate. With a goal of structurally characterizing the AT-ACP interface, a strategy was developed for covalently cross-linking the active site Ser → Cys mutant of the DSZS AT to its ACP substrate and for purifying the resulting AT-ACP complex to homogeneity. The S86C DSZS AT mutant was found to be functional, albeit with a transacylation efficiency 200-fold lower than that of its wild-type counterpart. Our findings provide new insights as well as new opportunities for high-resolution analysis of an important protein-protein interface in polyketide synthases.

PubMed: 21707057
DOI: 10.1021/bi200632j

Experimental method

X-RAY DIFFRACTION (1.35 Å)