Ultraviolet Absorption Spectroscopy of Peptides

Springer Protocols

(2014)

Authors:

Mangala R. Liyanage 1 ,

Kunal Bakshi 1 ,

David B. Volkin 1 ,

C. Russell Middaugh 1

Mangala R. Liyanage 1 ,

Kunal Bakshi 1 ,

David B. Volkin 1 ,

C. Russell Middaugh 1

Protocol | DOI: 10.1007/978-1-62703-673-3_15

Affiliations:

1. Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA

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Citations: 3

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Abstract

UV absorption spectroscopy is commonly used with peptides for determining concentration and enzyme activity, but high-resolution UV spectra can also provide information on peptide secondary and tertiary structure and association behavior. New

 

…more

UV absorption spectroscopy is commonly used with peptides for determining concentration and enzyme activity, but high-resolution UV spectra can also provide information on peptide secondary and tertiary structure and association behavior. New developments using temperature- and cation-dependent high-resolution second derivative absorption methods can also provide information concerning peptide dynamics. Data from several low-resolution spectroscopic techniques, including UV absorption, can be combined to generate an overall picture of peptide structure as a function of environmental conditions.

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Figures (0) & Videos (0)

• Fig. 1

• Fig. 1

Experimental Specifications

Techniques

Reagents

Other Keywords

Amino Acid Analysis

Structure Analysis

Ultraviolet Spectrophotometry

Protein Structure Analysis

Absorption Spectroscopy

Light Scattering

Circular Dichroism

Data analysis

Antibodies

Protein

Experimental Models

Kits

Please check back later as we are actively working on gathering the necessary information. Thank you for your understanding and patience.

Peptide

Stability

UV absorption

Ultraviolet

Near UV

Far UV

Conformational integrity

Techniques

Amino Acid Analysis

Structure Analysis

Ultraviolet Spectrophotometry

Protein Structure Analysis

Absorption Spectroscopy

Light Scattering

Circular Dichroism

Data analysis

Reagents

Please check back later as we are actively working on gathering the necessary information. Thank you for your understanding and patience.

Other Keywords

Peptide

Stability

UV absorption

Ultraviolet

Near UV

Far UV

Conformational integrity

Discover related articles

References

1. Pace CN, Vajdos F, Fee L, Grimsley G, Gray T (1995) How to measure and predict the molar absorption coefficient of a protein. Protein Sci 4:2411–2423
2. Mach H, Middaugh CR, Lewis RV (1992) Statistical determination of the average values of the extinction coefficients of tryptophan and tyrosine in native proteins. Anal Biochem 200(1):74–80
3. Edelhoch H (1967) Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry 6:1948–1954
4. Show more references

1. Pace CN, Vajdos F, Fee L, Grimsley G, Gray T (1995) How to measure and predict the molar absorption coefficient of a protein. Protein Sci 4:2411–2423
2. Mach H, Middaugh CR, Lewis RV (1992) Statistical determination of the average values of the extinction coefficients of tryptophan and tyrosine in native proteins. Anal Biochem 200(1):74–80
3. Edelhoch H (1967) Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry 6:1948–1954
4. Gill SC, von Hippel PH (1989) Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem 182:319–326
5. Weiser WE, Pardue HL (1983) Evaluation of multi-wavelength derivative spectra for quantitative applications in clinical chemistry. Clin Chem 29(9):1673–1677
6. Mach H, Middaugh CR (1993) Measuring protein spectra in the presence of light scattering. Biotechniques 15(2):240–242
7. Lucas LH, Ersoy BA, Kueltzo LA, Joshi SB, Brandau DT, Thyagarajapuram N, Peek LJ, Middaugh CR (2006) Probing protein structure and dynamics by second-derivative ultraviolet absorption analysis of cation-π interactions. Protein Sci 15(10):2228–2243
8. Esfandiary R, Hunjan JS, Lushington GH, Joshi SB, Middaugh CR (2009) Temperature dependent 2nd derivative absorbance spectroscopy of aromatic amino acids as a probe of protein dynamics. Protein Sci 18(12):2603–2614
9. Wegner A, Savko P (1982) Fragmentation of actin filaments. Biochemistry 21(8):1909–1913
10. Hofrichter J, Ross PD, Eaton WA (1976) Supersaturation in sickle cell hemoglobin solutions. Proc Natl Acad Sci U S A 73(9):3035–3039
11. Tomski SJ, Murphy RM (1992) Kinetics of aggregation of synthetic beta-amyloid peptide. Arch Biochem Biophys 294:630–638
12. Snyder SW, Ladror US, Wade WS, Wang GT, Barrett LW (1994) Amyloid-beta aggregation: selective inhibition of aggregation in mixtures of amyloid with different chain lengths. Biophys J 67:1216–1228
13. Schneider AS (1973) Analysis of optical activity spectra in turbid biological suspensions. Methods Enzymol 27:751–767
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Abstract

UV absorption spectroscopy is commonly used with peptides for determining concentration and enzyme activity, but high-resolution UV spectra can also provide information on peptide secondary and tertiary structure and association behavior. New

 

…more

UV absorption spectroscopy is commonly used with peptides for determining concentration and enzyme activity, but high-resolution UV spectra can also provide information on peptide secondary and tertiary structure and association behavior. New developments using temperature- and cation-dependent high-resolution second derivative absorption methods can also provide information concerning peptide dynamics. Data from several low-resolution spectroscopic techniques, including UV absorption, can be combined to generate an overall picture of peptide structure as a function of environmental conditions.

less

Experimental Specifications

Techniques

Reagents

Other Keywords

Amino Acid Analysis

Structure Analysis

Ultraviolet Spectrophotometry

Protein Structure Analysis

Absorption Spectroscopy

Light Scattering

Circular Dichroism

Data analysis

Antibodies

Protein

Experimental Models

Kits

Please check back later as we are actively working on gathering the necessary information. Thank you for your understanding and patience.

Peptide

Stability

UV absorption

Ultraviolet

Near UV

Far UV

Conformational integrity

Techniques

Amino Acid Analysis

Structure Analysis

Ultraviolet Spectrophotometry

Protein Structure Analysis

Absorption Spectroscopy

Light Scattering

Circular Dichroism

Data analysis

Reagents

Please check back later as we are actively working on gathering the necessary information. Thank you for your understanding and patience.

Other Keywords

Peptide

Stability

UV absorption

Ultraviolet

Near UV

Far UV

Conformational integrity

Discover related articles

References

1. Pace CN, Vajdos F, Fee L, Grimsley G, Gray T (1995) How to measure and predict the molar absorption coefficient of a protein. Protein Sci 4:2411–2423
2. Mach H, Middaugh CR, Lewis RV (1992) Statistical determination of the average values of the extinction coefficients of tryptophan and tyrosine in native proteins. Anal Biochem 200(1):74–80
3. Edelhoch H (1967) Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry 6:1948–1954
4. Show more references

1. Pace CN, Vajdos F, Fee L, Grimsley G, Gray T (1995) How to measure and predict the molar absorption coefficient of a protein. Protein Sci 4:2411–2423
2. Mach H, Middaugh CR, Lewis RV (1992) Statistical determination of the average values of the extinction coefficients of tryptophan and tyrosine in native proteins. Anal Biochem 200(1):74–80
3. Edelhoch H (1967) Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry 6:1948–1954
4. Gill SC, von Hippel PH (1989) Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem 182:319–326
5. Weiser WE, Pardue HL (1983) Evaluation of multi-wavelength derivative spectra for quantitative applications in clinical chemistry. Clin Chem 29(9):1673–1677
6. Mach H, Middaugh CR (1993) Measuring protein spectra in the presence of light scattering. Biotechniques 15(2):240–242
7. Lucas LH, Ersoy BA, Kueltzo LA, Joshi SB, Brandau DT, Thyagarajapuram N, Peek LJ, Middaugh CR (2006) Probing protein structure and dynamics by second-derivative ultraviolet absorption analysis of cation-π interactions. Protein Sci 15(10):2228–2243
8. Esfandiary R, Hunjan JS, Lushington GH, Joshi SB, Middaugh CR (2009) Temperature dependent 2nd derivative absorbance spectroscopy of aromatic amino acids as a probe of protein dynamics. Protein Sci 18(12):2603–2614
9. Wegner A, Savko P (1982) Fragmentation of actin filaments. Biochemistry 21(8):1909–1913
10. Hofrichter J, Ross PD, Eaton WA (1976) Supersaturation in sickle cell hemoglobin solutions. Proc Natl Acad Sci U S A 73(9):3035–3039
11. Tomski SJ, Murphy RM (1992) Kinetics of aggregation of synthetic beta-amyloid peptide. Arch Biochem Biophys 294:630–638
12. Snyder SW, Ladror US, Wade WS, Wang GT, Barrett LW (1994) Amyloid-beta aggregation: selective inhibition of aggregation in mixtures of amyloid with different chain lengths. Biophys J 67:1216–1228
13. Schneider AS (1973) Analysis of optical activity spectra in turbid biological suspensions. Methods Enzymol 27:751–767
14. less

Figures (0) & Videos (0)

• Fig. 1

• Fig. 1