title: Molecular mechanisms of the anomalous thermal aggregation of green fluorescent protein.
abstract: The peculiarities of thermal denaturation and interaction with water of the cycle-3 mutant of green fluorescent protein (GFP) were analyzed by NMR techniques and compared with those of bovine carbonic anhydrase II (BCA-II). Irreversible thermal denaturation was accompanied by massive GFP aggregation with no detectable accumulation of soluble denatured protein. Analysis of the spin diffusion data suggested that the internal part of the GFP β-can is involved in intensive interactions with water molecules. As a result, at high temperatures, the GFP structure does not unfold but rather breaks, consequently leading to enhanced protein aggregation. This is very different from typical BCA-II behavior.
authors.affiliation: Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, Russia.
authors.forename: Bogan S
authors.initials: BS
authors.lastname: Melnik
authors.forename: Nikolay V
authors.initials: NV
authors.lastname: Molochkov
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Prokhorov
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Uversky
authors.forename: Viktor P
authors.initials: VP
authors.lastname: Kutyshenko
mesh_terms: D000818:Animals; D002417:Cattle; D011232:Chemical Precipitation; D004058:Diffusion; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding; D055503:Protein Multimerization / Q000187:drug effects; D012997:Solvents / Q000494:pharmacology; D013696:Temperature; D013816:Thermodynamics; D014867:Water / Q000737:chemistry / Q000378:metabolism
chemical_list: D012997:Solvents; D014867:Water; D049452:Green Fluorescent Proteins