The length of the bound fatty acid influences the dynamics of the acyl carrier protein and the stability of the thioester bond.

Acyl carrier proteins involved in fatty acid biosynthesis have been shown to exhibit a high degree of conformational flexibility, in that they are able to sequester fatty acid intermediates between 4 and 18 carbons in length. This flexibility has been observed in X-ray and NMR structures of acyl carrier proteins attached to different fatty acids. NMR studies comparing decanoyl-ACP and stearoyl-ACP indicated that ACP exhibits more dynamic motions when bound to longer fatty acids. We have used complementary chemical and NMR methods as an approach to improving our understanding of the effect of fatty acid length on the dynamics of acyl carrier protein. A chemical assay of the accessibility of the acyl thioester to solvent revealed a positive correlation between chain length and rate of hydrolysis. Surprisingly, this linear correlation was biphasic, with accelerated hydrolysis observed for fatty acids longer than 15 carbons. To further understand the motions associated with this acceleration, we collected (15)N relaxation dispersion data for 14:0-, 15:0-, and 16:0-ACP. The greatest dispersions were exhibited by residues that form the entrance to the fatty acid binding pocket. In addition, these dispersions were observed to increase with the length of the fatty acid. Because the exchange rates derived from fitting the data to a two-state model varied from residue to residue, a more complex motional model appears to be required to adequately explain the dynamics. Thus, acyl-ACP offers an interesting system for future investigations of complex protein motions on the micro- and millisecond time scales.