Abstract
Elucidating proteins function at a level that allows for intelligent design and manipulation is essential in realization of their potential role in biomedical and industrial applications. It has become increasingly apparent though, that probing structures and functionalities under equilibrium conditions is not sufficient. Rather, many aspects of protein behavior and reactivity are rooted in protein dynamics. Thus, there is a growing effort to probe intermediate structures that occur transiently during the course of a proteins function in particular linked to the binding or release of a ligand or substrate. However, studies following the sequence of conformational changes triggered by the binding of substrate/ligand and the concomitant change in functional properties are inherently difficult because often the diffusion times are of the order of conformational relaxation times. This chapter describes methodologies for generating resonance Raman spectra from transient forms of hemoglobin under conditions that allow for the systematic exploration of conformational relaxation and functionality. Special consideration is given to Raman compatible protocols based on sol-gel encapsulation that allow for the preparation, trapping and temporal tuning of nonequilibrium population generated from either the addition or the removal of ligands/substrates.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 287-300 |
| Number of pages | 14 |
| Journal | Methods in molecular biology (Clifton, N.J.) |
| Volume | 305 |
| State | Published - 2005 |
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ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Proteins in motion : resonance Raman spectroscopy as a probe of functional intermediates. / Samuni, Uri; Friedman, Joel M.
In: Methods in molecular biology (Clifton, N.J.), Vol. 305, 2005, p. 287-300.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Proteins in motion
T2 - resonance Raman spectroscopy as a probe of functional intermediates.
AU - Samuni, Uri
AU - Friedman, Joel M.
PY - 2005
Y1 - 2005
N2 - Elucidating proteins function at a level that allows for intelligent design and manipulation is essential in realization of their potential role in biomedical and industrial applications. It has become increasingly apparent though, that probing structures and functionalities under equilibrium conditions is not sufficient. Rather, many aspects of protein behavior and reactivity are rooted in protein dynamics. Thus, there is a growing effort to probe intermediate structures that occur transiently during the course of a proteins function in particular linked to the binding or release of a ligand or substrate. However, studies following the sequence of conformational changes triggered by the binding of substrate/ligand and the concomitant change in functional properties are inherently difficult because often the diffusion times are of the order of conformational relaxation times. This chapter describes methodologies for generating resonance Raman spectra from transient forms of hemoglobin under conditions that allow for the systematic exploration of conformational relaxation and functionality. Special consideration is given to Raman compatible protocols based on sol-gel encapsulation that allow for the preparation, trapping and temporal tuning of nonequilibrium population generated from either the addition or the removal of ligands/substrates.
AB - Elucidating proteins function at a level that allows for intelligent design and manipulation is essential in realization of their potential role in biomedical and industrial applications. It has become increasingly apparent though, that probing structures and functionalities under equilibrium conditions is not sufficient. Rather, many aspects of protein behavior and reactivity are rooted in protein dynamics. Thus, there is a growing effort to probe intermediate structures that occur transiently during the course of a proteins function in particular linked to the binding or release of a ligand or substrate. However, studies following the sequence of conformational changes triggered by the binding of substrate/ligand and the concomitant change in functional properties are inherently difficult because often the diffusion times are of the order of conformational relaxation times. This chapter describes methodologies for generating resonance Raman spectra from transient forms of hemoglobin under conditions that allow for the systematic exploration of conformational relaxation and functionality. Special consideration is given to Raman compatible protocols based on sol-gel encapsulation that allow for the preparation, trapping and temporal tuning of nonequilibrium population generated from either the addition or the removal of ligands/substrates.
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M3 - Article
C2 - 15940003
AN - SCOPUS:21344452300
VL - 305
SP - 287
EP - 300
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -