Biochemical Characterization and Crystal Structure of a Novel NAD+-Dependent Isocitrate Dehydrogenase from Phaeodactylum tricornutum.

The marine diatom Phaeodactylum tricornutum originated from a series of secondary symbiotic events and has been used as a model organism for studying diatom biology. A novel type II homodimeric isocitrate dehydrogenase from P. tricornutum (PtIDH1) was expressed, purified, and identified in detail through enzymatic characterization. Kinetic analysis showed that PtIDH1 is NAD+-dependent and has no detectable activity with NADP+. The catalytic efficiency of PtIDH1 for NAD+ is 0.16 µM-1·s-1 and 0.09 µM-1·s-1 in the presence of Mn2+ and Mg2+, respectively. Unlike other bacterial homodimeric NAD-IDHs, PtIDH1 activity was allosterically regulated by the isocitrate. Furthermore, the dimeric structure of PtIDH1 was determined at 2.8 Å resolution, and each subunit was resolved into four domains, similar to the eukaryotic homodimeric NADP-IDH in the type II subfamily. Interestingly, a unique and novel C-terminal EF-hand domain was first defined in PtIDH1. Deletion of this domain disrupted the intact dimeric structure and activity. Mutation of the four Ca2+-binding sites in the EF-hand significantly reduced the calcium tolerance of PtIDH1. Thus, we suggest that the EF-hand domain could be involved in the dimerization and Ca2+-coordination of PtIDH1. The current report, on the first structure of type II eukaryotic NAD-IDH, provides new information for further investigation of the evolution of the IDH family.

First on-sky demonstration of the piezoelectric adaptive secondary mirror.

We propose using a piezoelectric adaptive secondary mirror (PASM) in the medium-sized adaptive telescopes with a 2-4 m aperture for structure and control simplification by utilizing the piezoelectric actuators in contrast with the voice-coil adaptive secondary mirror. A closed-loop experimental setup was built for on-sky demonstration of the 73-element PASM developed by our laboratory. In this Letter, the PASM and the closed-loop adaptive optics system are introduced. High-resolution stellar images were obtained by using the PASM to correct high-order wavefront errors in May 2016. To the best of our knowledge, this is the first successful on-sky demonstration of the PASM. The results show that with the PASM as the deformable mirror, the angular resolution of the 1.8 m telescope can be effectively improved.