Time-resolved fluorescence. An approach in protein analysis.
This paper deal with the spectroscopic analysis of proteins, i.e., human serum albumin (HSA) and bovine serum albumin (BSA). These two proteins present very similar UV absorption and fluorescence spectra. The UV-spectra of both proteins result from the sum of the absorption spectra of the aromatic amino acid, mainly tryptophan and tyrosine, present in the biological molecules. On the contrary, the fluorescence HSA and BSA spectra are not the sum of the emission fluorescence spectra of tyrosine and tryptophan, but they are mainly constituted by the tryptophan fluorescence with a poor contribution arising from tyrosine. This fact may be due to inter- and intramolecular quenching phenomena. Significant difference were recorder in the life time fluorescence decay of HSA and BSA, which showed tau values of 2.3 and 4.5 nanoseconds, respectively. The time-resolved fluorescence spectroscopy was performed with an experimental apparatus assembled in our laboratories, which is able of measuring the fluorescence decay in the range 2 x 10(-10)-2 x 10(-8) sec.
Villari
A
A
Dipartimento Farmaco-Chimico, Facoltà di Farmacia, Università di Messina, Italy.
Micali
N
N
Fresta
M
M
Trusso
S
S
Puglisi
G
G
D000818:Animals; D002417:Cattle; D006801:Humans; D008962:Models, Theoretical; D010649:Phenylalanine / Q000032:analysis; D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013997:Time Factors; D014364:Tryptophan / Q000032:analysis; D014443:Tyrosine / Q000032:analysis
D011506:Proteins; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D014443:Tyrosine; D010649:Phenylalanine; D014364:Tryptophan