Squash glycerol-3-phospate-1-acyltransferase has been crystallized and the structure of the enzyme determined, at 1.9-Å resolution, using multiple isomorphous replacement of the wild type and a series of individual cysteine mutants. Competitive in vitro substrate selectivity assays have been established that differentiate between selective and non-selective forms of the enzyme. Particular care was taken to use near-physiological concentrations of both substrates. Clear substrate selectivity can be demonstrated with the natural substrate acyl-acyl carrier protein but not with the substrate analogue acyl-CoA. The use of site-directed mutagenesis, coupled to three-dimensional structural determinations, should provide a rational basis for elucidating structural components important in determining the substrate selectivity of this enzyme.
Plant glycerol-3-phosphate-l-acyltransferase (GPAT): structure selectivity studies
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A. R. Slabas, W. R. Simon, T. Schierer, J. Kroon, T. Fawcett, M. Hayman, J. Gilroy, I. Nishida, N. Murata, J. Rafferty, A. Turnbull, D. Rice; Plant glycerol-3-phosphate-l-acyltransferase (GPAT): structure selectivity studies. Biochem Soc Trans 1 December 2000; 28 (6): 677–679. doi: https://doi.org/10.1042/bst0280677
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