title: Identification of proteins interacting with lactate dehydrogenase in claw muscle of the porcelain crab Petrolisthes cinctipes.
abstract: Biochemical adaptation of enzymes involves conservation of activity, stability and affinity across a wide range of intracellular and environmental conditions. Enzyme adaptation by alteration of primary structure is well known, but the roles of protein-protein interactions in enzyme adaptation are less well understood. Interspecific differences in thermal stability of lactate dehydrogenase (LDH) in porcelain crabs (genus Petrolisthes) are related to intrinsic differences among LDH molecules and by interactions with other stabilizing proteins. Here, we identified proteins that interact with LDH in porcelain crab claw muscle tissue using co-immunoprecipitation, and showed LDH exists in high molecular weight complexes using size exclusion chromatography and Western blot analyses. Co-immunoprecipitated proteins were separated using 2D SDS PAGE and analyzed by LC/ESI using peptide MS/MS. Peptide MS/MS ions were compared to an EST database for Petrolisthes cinctipes to identify proteins. Identified proteins included cytoskeletal elements, glycolytic enzymes, a phosphagen kinase, and the respiratory protein hemocyanin. Our results support the hypothesis that LDH interacts with glycolytic enzymes in a metabolon structured by cytoskeletal elements that may also include the enzyme for transfer of the adenylate charge in glycolytically produced ATP. Those interactions may play specific roles in biochemical adaptation of glycolytic enzymes.
authors.affiliation: Romberg Tiburon Center and Department of Biology, San Francisco State University, 3152 Paradise Drive, Tiburon California 94920, USA.
authors.forename: Andrea P
authors.initials: AP
authors.lastname: Cayenne
authors.forename: Beverly
authors.initials: B
authors.lastname: Gabert
authors.forename: Jonathon H
authors.initials: JH
authors.lastname: Stillman
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000737:chemistry / Q000378:metabolism*; D015153:Blotting, Western; D002850:Chromatography, Gel; D033364:Decapoda / Q000737:chemistry / Q000201:enzymology*; D006019:Glycolysis; D047468:Immunoprecipitation; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000378:metabolism*; D009132:Muscles / Q000201:enzymology*; D025941:Protein Interaction Mapping; D053719:Tandem Mass Spectrometry
chemical_list: D060829:Arthropod Proteins; D007770:L-Lactate Dehydrogenase