Determinants of dual substrate specificity revealed by the crystal structure of homoisocitrate dehydrogenase from Thermus thermophilus in complex with homoisocitrate center dot Mg2+ center dot NADH

作者：Takahashi Kento; Tomita Takeo; Kuzuyama Tomohisa; Nishiyama Makoto^*

来源：Biochemical and Biophysical Research Communications, 2016, 478(4): 1688-1693.

DOI：10.1016/j.bbrc.2016.09.004

• beta-decarboxylating dehydrogenase
• Homoisocitrate dehydrogenase
• Crystal structure
• Thermus thermophilus

HICDH (Homoisocitrate dehydrogenase) is a member of the beta-decarboxylating dehydrogenase family that catalyzes the conversion of homoisocitrate to alpha-ketoadipate using NAD(+) as a coenzyme, which is the fourth reaction involved in lysine biosynthesis through the alpha-aminoadipate pathway. Although typical HICDHs from fungi and yeast exhibit strict substrate specificities toward homoisocitrate (HIC), HICDH from a thermophilic bacterium Thermus thermophilus (TtHICDH) catalyzes the reactions using both HIC and isocitrate (IC) as substrates at similar efficiencies. We herein determined the crystal structure of the quaternary complex of TtHICDH with HIC, NADH, and Mg2+ ion at a resolution of 2.5 angstrom. The structure revealed that the distal carboxyl group of HIC was recognized by the side chains of Ser72 and Arg85 from one subunit, and Asn173 from another subunit of a dimer unit. Model structures were constructed for TtHICDH in complex with IC and also for HICDH from Saccharomyces cerevisiae (ScHICDH) in complex with HIC. TtHICDH recognized the distal carboxyl group of IC by Arg85 in the model. In ScHICDH, the distal carboxyl group of HIC was recognized by the side chains of Ser98 and Ser108 from one subunit and Asn208 from another subunit of a dimer unit. By contrast, in ScHICDH, which lacks an Arg residue at the position corresponding to Arg85 in TtHICDH, these residues may not interact with the distal carboxyl group of shorter IC. These results provide a molecular basis for the differences in substrate specificities between TtHICDH and ScHICDH.

• 出版日期2016-9-30