Fluorescence based assessment of SDS induced hydrophobic collapse in globular proteins

S. Manjunath, Venkata Krishna Kanth Makani, Kapaettu Satyamoorthy, Bola Sadashiva Satish Rao, Gopalkrishna Bhat, Akriti Baby Kanth, Krishna Kishore Mahato

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

Abstract

The molecular mechanism of interaction between SDS and proteins is not clearly understood so far. According to the current knowledge SDS is known to interact with the hydrophobic regions of the proteins. Tryptophan and tyrosine are hydrophobic and hydrophilic aromatic amino acids respectively, which are also known for their intrinsic fluorescence nature in proteins. By observing the autofluorescence of both these hydrophobic and hydrophilic amino acids upon SDS treatment, information about SDS-protein interactions could be obtained. In the present study we have recorded the autofluorescence spectra of five globular proteins [Bovine serum albumin (BSA), Human serum albumin (HSA), Ribonuclease A (RNase A), Lysozyme and Trypsin] by the sequential excitation from 260nm to 295nm at every 5nm intervals. The results obtained clearly indicated BSA and HSA undergone hydrophobic collapse around their tryptophan moieties due to the increased folding of their secondary and tertiary structures upon SDS treatment. Trypsin on the other hand showed complete unfolding upon treatment with SDS. Lysozyme and RNase A did not show any difference in their autofluorescence upon SDS treatment may be due to the stability and fluorophores composition in them. The above results obtained with specific UV excitations clearly shown the tertiary folding and ensembles of the secondary and tertiary structures upon SDS treatment is governed by their stability and bonds stabilizing the proteins.

Original languageEnglish
Title of host publicationBiophysics, Biology, and Biophotonics
Subtitle of host publicationThe Crossroads
PublisherSPIE
Volume9719
ISBN (Electronic)9781628419535
DOIs
Publication statusPublished - 2016
EventBiophysics, Biology, and Biophotonics: The Crossroads - San Francisco, United States
Duration: 13-02-201614-02-2016

Conference

ConferenceBiophysics, Biology, and Biophotonics: The Crossroads
CountryUnited States
CitySan Francisco
Period13-02-1614-02-16

Fingerprint

Fluorescence
proteins
Proteins
albumins
serums
fluorescence
trypsin
Pancreatic Ribonuclease
tryptophan
lysozyme
Muramidase
Bovine Serum Albumin
Serum Albumin
Tryptophan
Trypsin
folding
amino acids
Amino acids
Enzymes
Aromatic Amino Acids

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., Makani, V. K. K., Satyamoorthy, K., Rao, B. S. S., Bhat, G., Kanth, A. B., & Mahato, K. K. (2016). Fluorescence based assessment of SDS induced hydrophobic collapse in globular proteins. In Biophysics, Biology, and Biophotonics: The Crossroads (Vol. 9719). [97190A] SPIE. https://doi.org/10.1117/12.2212708
Manjunath, S. ; Makani, Venkata Krishna Kanth ; Satyamoorthy, Kapaettu ; Rao, Bola Sadashiva Satish ; Bhat, Gopalkrishna ; Kanth, Akriti Baby ; Mahato, Krishna Kishore. / Fluorescence based assessment of SDS induced hydrophobic collapse in globular proteins. Biophysics, Biology, and Biophotonics: The Crossroads. Vol. 9719 SPIE, 2016.
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abstract = "The molecular mechanism of interaction between SDS and proteins is not clearly understood so far. According to the current knowledge SDS is known to interact with the hydrophobic regions of the proteins. Tryptophan and tyrosine are hydrophobic and hydrophilic aromatic amino acids respectively, which are also known for their intrinsic fluorescence nature in proteins. By observing the autofluorescence of both these hydrophobic and hydrophilic amino acids upon SDS treatment, information about SDS-protein interactions could be obtained. In the present study we have recorded the autofluorescence spectra of five globular proteins [Bovine serum albumin (BSA), Human serum albumin (HSA), Ribonuclease A (RNase A), Lysozyme and Trypsin] by the sequential excitation from 260nm to 295nm at every 5nm intervals. The results obtained clearly indicated BSA and HSA undergone hydrophobic collapse around their tryptophan moieties due to the increased folding of their secondary and tertiary structures upon SDS treatment. Trypsin on the other hand showed complete unfolding upon treatment with SDS. Lysozyme and RNase A did not show any difference in their autofluorescence upon SDS treatment may be due to the stability and fluorophores composition in them. The above results obtained with specific UV excitations clearly shown the tertiary folding and ensembles of the secondary and tertiary structures upon SDS treatment is governed by their stability and bonds stabilizing the proteins.",
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Manjunath, S, Makani, VKK, Satyamoorthy, K, Rao, BSS, Bhat, G, Kanth, AB & Mahato, KK 2016, Fluorescence based assessment of SDS induced hydrophobic collapse in globular proteins. in Biophysics, Biology, and Biophotonics: The Crossroads. vol. 9719, 97190A, SPIE, Biophysics, Biology, and Biophotonics: The Crossroads, San Francisco, United States, 13-02-16. https://doi.org/10.1117/12.2212708

Fluorescence based assessment of SDS induced hydrophobic collapse in globular proteins. / Manjunath, S.; Makani, Venkata Krishna Kanth; Satyamoorthy, Kapaettu; Rao, Bola Sadashiva Satish; Bhat, Gopalkrishna; Kanth, Akriti Baby; Mahato, Krishna Kishore.

Biophysics, Biology, and Biophotonics: The Crossroads. Vol. 9719 SPIE, 2016. 97190A.

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

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AU - Makani, Venkata Krishna Kanth

AU - Satyamoorthy, Kapaettu

AU - Rao, Bola Sadashiva Satish

AU - Bhat, Gopalkrishna

AU - Kanth, Akriti Baby

AU - Mahato, Krishna Kishore

PY - 2016

Y1 - 2016

N2 - The molecular mechanism of interaction between SDS and proteins is not clearly understood so far. According to the current knowledge SDS is known to interact with the hydrophobic regions of the proteins. Tryptophan and tyrosine are hydrophobic and hydrophilic aromatic amino acids respectively, which are also known for their intrinsic fluorescence nature in proteins. By observing the autofluorescence of both these hydrophobic and hydrophilic amino acids upon SDS treatment, information about SDS-protein interactions could be obtained. In the present study we have recorded the autofluorescence spectra of five globular proteins [Bovine serum albumin (BSA), Human serum albumin (HSA), Ribonuclease A (RNase A), Lysozyme and Trypsin] by the sequential excitation from 260nm to 295nm at every 5nm intervals. The results obtained clearly indicated BSA and HSA undergone hydrophobic collapse around their tryptophan moieties due to the increased folding of their secondary and tertiary structures upon SDS treatment. Trypsin on the other hand showed complete unfolding upon treatment with SDS. Lysozyme and RNase A did not show any difference in their autofluorescence upon SDS treatment may be due to the stability and fluorophores composition in them. The above results obtained with specific UV excitations clearly shown the tertiary folding and ensembles of the secondary and tertiary structures upon SDS treatment is governed by their stability and bonds stabilizing the proteins.

AB - The molecular mechanism of interaction between SDS and proteins is not clearly understood so far. According to the current knowledge SDS is known to interact with the hydrophobic regions of the proteins. Tryptophan and tyrosine are hydrophobic and hydrophilic aromatic amino acids respectively, which are also known for their intrinsic fluorescence nature in proteins. By observing the autofluorescence of both these hydrophobic and hydrophilic amino acids upon SDS treatment, information about SDS-protein interactions could be obtained. In the present study we have recorded the autofluorescence spectra of five globular proteins [Bovine serum albumin (BSA), Human serum albumin (HSA), Ribonuclease A (RNase A), Lysozyme and Trypsin] by the sequential excitation from 260nm to 295nm at every 5nm intervals. The results obtained clearly indicated BSA and HSA undergone hydrophobic collapse around their tryptophan moieties due to the increased folding of their secondary and tertiary structures upon SDS treatment. Trypsin on the other hand showed complete unfolding upon treatment with SDS. Lysozyme and RNase A did not show any difference in their autofluorescence upon SDS treatment may be due to the stability and fluorophores composition in them. The above results obtained with specific UV excitations clearly shown the tertiary folding and ensembles of the secondary and tertiary structures upon SDS treatment is governed by their stability and bonds stabilizing the proteins.

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PB - SPIE

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Manjunath S, Makani VKK, Satyamoorthy K, Rao BSS, Bhat G, Kanth AB et al. Fluorescence based assessment of SDS induced hydrophobic collapse in globular proteins. In Biophysics, Biology, and Biophotonics: The Crossroads. Vol. 9719. SPIE. 2016. 97190A https://doi.org/10.1117/12.2212708