Journal
BIOCHEMISTRY
Volume 50, Issue 30, Pages 6539-6548Publisher
AMER CHEMICAL SOC
DOI: 10.1021/bi200632j
Keywords
-
Categories
Funding
- National Institutes of Health [GM087934, GM022172]
- Agency of Science, Technology and Research (A*STAR), Singapore
- DOE Office of Basic Energy Sciences, Office of Biological and Environmental Research
- National Institutes of Health, National Center for Research Resources
- National Institute of General Medical Sciences
Ask authors/readers for more resources
The 1.51 angstrom 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 angstrom 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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available