Fluorescence characterization of type I collagen from normal and silicotic rats and its quenching dynamics induced by hypocrellin B.
In this paper, we studied the quenching mechanism of intrinsic fluorescence of type I collagen by a new type photosensitizer and fluorescence quencher, hypocrellin B (HB). It was indicated that type I collagen can emit Tyr-intrinsic fluorescence with the excitation wavelength of Tyr (lambda(ex) = 269 nm). Its fluorescence decay conform to the triexponential rule of the fluorescence lifetime. The intrinsic fluorescence of type I collagen can be effectively quenched by HB through a process of charge and energy transference, which is involved in the collisional quenching, the dipolar inducement, and the formation of exciplex between HB and excited fluorophores of collagen. The fluorescence quenching would be weakened by higher ionic environments. The fluorescence emission and its quenching rate of abnormal silicotic collagen show falling trends, implying its much weakened potential of charge and energy transference, and its lessen bioelectric activities. In conclusion, the bioelectric properties of collagen depends on the perfect order of its molecular structure and orderly intramolecular and intermolecular interactions, which is important in its performing normal physiological functions. It is also demonstrated that the fluorescence quenching technique, using HB as a quencher, is truly an effectively method for biomolecular studies.
Li
Y H
YH
Department of Biological Science and Biotechnology, Tsinghua University, Beijing, P.R. China.
Yue
J C
JC
Cai
G P
GP
D000818:Animals; D003094:Collagen / Q000096:biosynthesis / Q000737:chemistry* / Q000187:drug effects; D008168:Lung / Q000378:metabolism*; D010569:Perylene / Q000031:analogs & derivatives* / Q000494:pharmacology; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011487:Protein Conformation; D011809:Quinones / Q000494:pharmacology*; D051381:Rats; D012829:Silicosis / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
D017319:Photosensitizing Agents; D011809:Quinones; D010569:Perylene; D003094:Collagen; C064522:hypocrellin B