This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Lactate dehydrogenase (LDH) from chicken breast muscle is an essential enzyme in carbohydrate metabolism. Although amino acid data and atomic coordinate structure information are available for orthologous homologs, the structure of chicken muscle LDH-A had not yet been determined. LDH-A from chicken breast muscle was isolated and purified. Native chicken LDH-A and various substrate, cofactor and inhibitor complexes were crystallized. Crystals of native protein, as well as pyruvate, NAD, NADH and oxalate complexes, were flash cooled and shipped to SSRL, where diffraction data were collected on beam lines BL9-1 and BL9-2. Some data was collected remotely from CSU Fullerton as part of a Macromolecular Crystallography course. The native chicken structure (a=84.04 b=126.78 c=252.74 has been solved to 1.9 resolution by molecular replacement, using porcine LDH-A as the search model. The structure of LDH-A complexed with pyruvate (space group C2, a=75.16 b=152.40 c=142.79 beta=93.5 ) has been also been solved to 1.9 resolution by molecular replacement, using the native chicken structure as the search model, with one tetramer in the asymmetric unit. Structure determination and refinement have begun using 2.3 and 1.9 data for LDH-A complexed with NAD and NADH respectively. New crystals of chicken LDH-A, in complex with lactate and oxalate, have been grown.
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