title: Reduced transglutaminase-catalyzed protein aggregation is observed in the presence of creatine using sedimentation velocity.
abstract: Transglutaminases (TGases) are enzymes that catalyze covalent isopeptide crosslinks between reactive lysine and glutamine residues in proteins. Higher than normal local concentrations of TGase have been correlated with increased protein aggregation in vivo. These insoluble protein aggregates are the hallmark of several neurodegenerative diseases, including Alzheimer's, Parkinson's, and Huntington's diseases, although each aggregating protein involved is disease specific. Because TGase is implicated in protein aggregation, there is evidence that its regulation may retard disease progression. Here we report on a laser light transmission technique as an in vitro tool to gauge the efficacy of creatine, a candidate inhibitor, to regulate aggregation. Sedimentation velocities of protein-coated particles in TGase-containing water-glycerol solutions were tracked with different levels of creatine. Sedimentation velocities were converted to apparent aggregate sizes using Stoke's law of sedimentation. The results indicated that creatine promoted up to a 20% reduction in protein aggregation in vitro. This technique may prove to be useful in identifying other functional TGase inhibitors.
authors.affiliation: Department of Materials Science and Engineering, Virginia Polytechnic Institute and State University, Blacksburg, 24060, USA.
authors.forename: Elena F
authors.initials: EF
authors.lastname: Burguera
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Love
mesh_terms: D002499:Centrifugation, Density Gradient; D003401:Creatine / Q000494:pharmacology*; D006801:Humans; D007834:Lasers; D008863:Microspheres; D019636:Neurodegenerative Diseases / Q000503:physiopathology*; D011137:Polystyrenes; D011485:Protein Binding / Q000187:drug effects; D020836:Protein Structure, Quaternary; D012542:Scattering, Radiation; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014783:Viscosity
chemical_list: D011137:Polystyrenes; D011503:Transglutaminases; D003401:Creatine