27 February 2014 Nature of autofluorescence in human serum albumin under its native, unfolding and digested forms
Author Affiliations +
Abstract
Autofluorescence characteristics of human serum albumin (HSA) are highly sensitive to its local environment. Identification and characterization of the proteins in normal and disease conditions may have great clinical implications. Aim of the present study was to understand how autofluorescence properties of HSA varies with denaturation under urea (3.0M, 6.0M, 9.0M) and guanidine hydrochloride (GnHCl) (2.0M, 4.0M, 6.0M) as well as digestion with trypsin. Towards this, we have recorded the corresponding autofluorescence spectra of HSA at 281nm laser excitation and compared the outcomes. Although, HSA contains 1 tryptophan and 17 tyrosine residues, it has shown intense autofluorescence due to tryptophan as compared to the tyrosine in native form, which may be due to the fluorescence resonance energy transfer (FRET) from tyrosine to tryptophan. As the unfolding progresses in denatured and digested forms of the protein, a clear increase in tyrosine fluorescence as compared to tryptophan was observed, which may be due to the increase of tryptophan - tyrosine separation disturbing the FRET between them resulting in differences in the overall autofluorescence properties. The decrease in tryptophan fluorescence of around 17% in urea denatured, 32% in GnHCl denatured and 96% in tryptic digested HSA was observed as compared to its native form. The obtained results show a clear decrease in FRET between tyrosine and tryptophan residues with the progression of unfolding and urea seems to be less efficient than GnHCl in unfolding of HSA. These results demonstrate the potential of autofluorescence in characterizing proteins in general and HSA in particular.
© (2014) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Manjunath S., Manjunath S., Bola Sadashiva Satish Rao, Bola Sadashiva Satish Rao, Kapaettu Satyamoorthy, Kapaettu Satyamoorthy, Krishna Kishore Mahato, Krishna Kishore Mahato, } "Nature of autofluorescence in human serum albumin under its native, unfolding and digested forms", Proc. SPIE 8935, Advanced Biomedical and Clinical Diagnostic Systems XII, 893520 (27 February 2014); doi: 10.1117/12.2039767; https://doi.org/10.1117/12.2039767
PROCEEDINGS
6 PAGES


SHARE
Back to Top