[Effect of calcium binding on the quenching time of self-fluorescence of Ca-binding proteins].
Lifetimes of phenylalanine, tyrosine and tryptophan self-fluorescence of three Ca2+-binding proteins (parvalbumins pI 4.47 and 3.95 and bovine alpha-lactalbumin) in the Ca2+-saturated state and without Ca2+ were measured on a device functioning in a channel of synchrotron radiation of the Lebedev Physical Institute electron accelerator C-60 with a single photon counting system. The decay curve of phenylalanine fluorescence of Ca2+-saturated parvalbumin pI 4.47 is two-exponential, which results from the presence of two subsystems of phenylalanine residues in this protein. Radiation of these subsystems is almost independent of one another. Detachment of Ca2+ from protein disturbs these subsystems. In case of tyrosine fluorescence of carp parvalbumin pI 3.95 a change in the quantum yield value of the stationary fluorescence induced by elimination of Ca2+ proceeds without a change of fluorescence lifetime. This seems to be related to the existence of static quenching of fluorescence in this case at the expense of complex formation between the chromophore and some adjacent quenching groups. Detachment of Ca2+ from alpha-lactalbumin induces conformational changes in its structure. The latter result in a transition of a number of tryptophane residues from its interior to the surface of the globule which is reflected in an increase of fluorescence quantum yield duration. It is concluded that in Ca2+-saturated alpha-lactalbumin some tryptophane residues are located near the quenching groups (dynamic quenching), most likely the disulfide bridges.
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002417:Cattle; D007700:Kinetics; D007768:Lactalbumin / Q000378:metabolism*; D009124:Muscle Proteins / Q000378:metabolism*; D010320:Parvalbumins / Q000378:metabolism*; D010649:Phenylalanine / Q000032:analysis; D011485:Protein Binding; D013050:Spectrometry, Fluorescence
D009124:Muscle Proteins; D010320:Parvalbumins; D010649:Phenylalanine; D007768:Lactalbumin; D002118:Calcium
Vliianie sviazyvaniia kal'tsiia na vremia zatukhaniia sobstvennoĭ fluorestsentsii Ca-sviazyvaiushchikh belkov.