title: Effects of fluorescent pseudo-ATP and ATP-metal analogs on secondary structure of Na(+)/K(+)-ATPase.
abstract: The secondary structure of Na(+)/K(+)-ATPase after modification of the ATP-binding sites was analyzed. Consistently with recent reports, we found in trypsin-treated Na(+)/K(+)-ATPase additionally to alpha-helix also beta-sheet structures in the transmembrane segments. However, binding of fluorescein 5'-isothiocyanate (FITC), the pseudo-ATP analog, to the ATP-binding site did not affect the secondary structure of undigested Na(+)/K(+)-ATPase. Consequently, fluorescence intensity changes of FITC-labeled Na(+)/K(+)-ATPase commonly used to observe conformational transitions of the enzyme reflect physiological changes of the native structure. The metal complex analogues of ATP, Cr(H(2)O)(4)ATP and Co(NH(3))(4)ATP, on the other hand, affected the secondary structure of Na(+)/K(+)-ATPase. We propose that these changes in the secondary structure are responsible for inhibition of backdoor phosphorylation.
authors.affiliation: Institute of Biochemistry, Medical School, University of Ancona, I-60131, Ancona, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Tanfani
authors.forename: H
authors.initials: H
authors.lastname: Linnertz
authors.forename: T
authors.initials: T
authors.lastname: Obsil
authors.forename: R
authors.initials: R
authors.lastname: Krumscheid
authors.forename: P
authors.initials: P
authors.lastname: Urbanova
authors.forename: O
authors.initials: O
authors.lastname: Jelinek
authors.forename: L
authors.initials: L
authors.lastname: Mazzanti
authors.forename: E
authors.initials: E
authors.lastname: Bertoli
authors.forename: W
authors.initials: W
authors.lastname: Schoner
authors.forename: E
authors.initials: E
authors.lastname: Amler
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives; D004923:Erythrosine / Q000031:analogs & derivatives; D016650:Fluorescein-5-isothiocyanate; D017879:Isothiocyanates; D009942:Organometallic Compounds; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared; D014364:Tryptophan / Q000737:chemistry
chemical_list: D017879:Isothiocyanates; D009942:Organometallic Compounds; C020511:tetraamminecobalt(III)ATP; C022232:erythrosine isothiocyanate; D014364:Tryptophan; D000255:Adenosine Triphosphate; D000254:Sodium-Potassium-Exchanging ATPase; D016650:Fluorescein-5-isothiocyanate; D004923:Erythrosine