Nature of autofluorescence in human serum albumin under its native, unfolding and digested forms

S. Manjunath, Bola Sadashiva Satish Rao, Kapaettu Satyamoorthy, Krishna Kishore Mahato

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

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.

Original languageEnglish
Title of host publicationAdvanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII
PublisherSPIE
Volume8935
ISBN (Print)9780819498489
DOIs
Publication statusPublished - 2014
EventAdvanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII - San Francisco, CA, United States
Duration: 02-02-201404-02-2014

Conference

ConferenceAdvanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII
CountryUnited States
CitySan Francisco, CA
Period02-02-1404-02-14

Fingerprint

tryptophan
albumins
tyrosine
Urea
Serum Albumin
Tryptophan
serums
Tyrosine
Proteins
guanidines
Fluorescence Resonance Energy Transfer
Fluorescence
Guanidine
hydrochlorides
resonance fluorescence
ureas
Denaturation
Laser excitation
energy transfer
proteins

All Science Journal Classification (ASJC) codes

  • Atomic and Molecular Physics, and Optics
  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

Cite this

Manjunath, S., Rao, B. S. S., Satyamoorthy, K., & Mahato, K. K. (2014). Nature of autofluorescence in human serum albumin under its native, unfolding and digested forms. In Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII (Vol. 8935). [893520] SPIE. https://doi.org/10.1117/12.2039767
Manjunath, S. ; Rao, Bola Sadashiva Satish ; Satyamoorthy, Kapaettu ; Mahato, Krishna Kishore. / Nature of autofluorescence in human serum albumin under its native, unfolding and digested forms. Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII. Vol. 8935 SPIE, 2014.
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Manjunath, S, Rao, BSS, Satyamoorthy, K & Mahato, KK 2014, Nature of autofluorescence in human serum albumin under its native, unfolding and digested forms. in Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII. vol. 8935, 893520, SPIE, Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII, San Francisco, CA, United States, 02-02-14. https://doi.org/10.1117/12.2039767

Nature of autofluorescence in human serum albumin under its native, unfolding and digested forms. / Manjunath, S.; Rao, Bola Sadashiva Satish; Satyamoorthy, Kapaettu; Mahato, Krishna Kishore.

Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII. Vol. 8935 SPIE, 2014. 893520.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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AB - 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.

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Manjunath S, Rao BSS, Satyamoorthy K, Mahato KK. Nature of autofluorescence in human serum albumin under its native, unfolding and digested forms. In Advanced Biomedical and Clinical Diagnostic and Surgical Guidance Systems XII. Vol. 8935. SPIE. 2014. 893520 https://doi.org/10.1117/12.2039767