title: Influence of ADP, AMP-PNP and of depletion of nucleotides on the structural properties of F1ATPase: a Fourier transform infrared spectroscopic study.
abstract: Mitochondrial F1ATPase from beef heart was treated with different buffers in order to modulate the nucleotide content of the enzyme and then analysed by FT-IR spectroscopy. Treatment of F1ATPase with a buffer lacking nucleotides and glycerol led to the formation of two fractions consisting of an inactive aggregated enzyme deprived almost completely of bound nucleotides and of an active enzyme containing ATP only in the tight sites and having a structure largely accessible to the solvent and a low thermal stability. Treatment of F1ATPase with saturating ADP, which induced the hysteretic inhibition during turnover, or AMP-PNP did not affect remarkably the secondary structure of the enzyme complex but significantly increased its compactness and thermal stability. It was hypothesised that the formation of the inactive aggregated enzyme was mainly due to the destabilisation of the alpha-subunits of F1ATPase and that the induction of the hysteretic inhibition is related to a particular conformation of the enzyme, which during turnover becomes unable to sustain catalysis.
authors.affiliation: Dipartimento di Scienze e Tecnologie Biomediche, Università di Udine, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Lippe
authors.forename: F
authors.initials: F
authors.lastname: Di Pancrazio
authors.forename: F
authors.initials: F
authors.lastname: Dabbeni-Sala
authors.forename: E
authors.initials: E
authors.lastname: Bertoli
authors.forename: F
authors.initials: F
authors.lastname: Tanfani
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology*; D000266:Adenylyl Imidodiphosphate / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D008929:Mitochondria, Heart / Q000201:enzymology; D009711:Nucleotides / Q000378:metabolism* / Q000494:pharmacology; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000187:drug effects* / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods; D013816:Thermodynamics
chemical_list: D009711:Nucleotides; D000266:Adenylyl Imidodiphosphate; D000244:Adenosine Diphosphate; D006180:Proton-Translocating ATPases