Document Detail


A strongly absorbing class of non-natural labels for probing protein electrostatics and solvation with FTIR and 2D IR spectroscopies.
MedLine Citation:
PMID:  23537223     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A series of non-natural infrared probes is reported that consist of a metal-tricarbonyl modified with a -(CH2)n- linker and cysteine-specific leaving group. They can be site-specifically attached to proteins using mutagenesis and similar protocols for EPR spin labels, which have the same leaving group. We characterize the label's frequencies and lifetimes using 2D IR spectroscopy in solvents of varying dielectric. The frequency range spans 10 cm(-1), and the variation in lifetimes ranges from 6 to 19 ps, indicating that these probes are very sensitive to their environments. Also, we attached probes with -(CH2)-, -(CH2)3-, and -(CH2)4- linkers to ubiquitin at positions 6 and 63 and collected spectra in aqueous buffer. The frequencies and lifetimes were correlated for 3C and 4C linkers, as they were in the solvents, but did not correlate for the 1C linker. We conclude that lifetime measures solvation, whereas frequency reflects the electrostatics of the environment, which in the case of the 1C linker is a measure of the protein electrostatic field. We also labeled V71C α-synuclein in buffer and membrane-bound. Unlike most other infrared labels, this label has extremely strong cross sections and thus can be measured with 2D IR spectroscopy at sub-millimolar concentrations. We expect that these labels will find use in studying the structure and dynamics of membrane-bound, aggregated, and kinetically evolving proteins for which high signal-to-noise at low protein concentrations is imperative.
Authors:
Ann Marie Woys; Sudipta S Mukherjee; David R Skoff; Sean D Moran; Martin T Zanni
Related Documents :
23839273 - Protein analysis by time-resolved measurements with an electro-switchable dna chip.
2821283 - Transcriptional and post-transcriptional controls establish the cascade of herpes simpl...
7717993 - Cyclophilin-40: evidence for a dimeric complex with hsp90.
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2013-04-15
Journal Detail:
Title:  The journal of physical chemistry. B     Volume:  117     ISSN:  1520-5207     ISO Abbreviation:  J Phys Chem B     Publication Date:  2013 May 
Date Detail:
Created Date:  2013-05-02     Completed Date:  2013-11-26     Revised Date:  2014-05-07    
Medline Journal Info:
Nlm Unique ID:  101157530     Medline TA:  J Phys Chem B     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5009-18     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Electron Spin Resonance Spectroscopy
Mesylates / chemistry
Solvents / chemistry*
Spectroscopy, Fourier Transform Infrared*
Spin Labels
Static Electricity
Ubiquitin / chemistry,  metabolism
alpha-Synuclein / chemistry*,  metabolism
Grant Support
ID/Acronym/Agency:
1 S10 RR13866-01/RR/NCRR NIH HHS; DK79895/DK/NIDDK NIH HHS; P41-GM103311/GM/NIGMS NIH HHS; P41GM66326/GM/NIGMS NIH HHS; P41RR02301/RR/NCRR NIH HHS; R01 DK079895/DK/NIDDK NIH HHS; RR02781/RR/NCRR NIH HHS; RR08438/RR/NCRR NIH HHS; T32 GM08293/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Mesylates; 0/Solvents; 0/Spin Labels; 0/Ubiquitin; 0/alpha-Synuclein; 44059-82-7/methanethiosulfonate
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Relationship between the population incidence of pertussis in children in New South Wales, Australia...
Next Document:  The effect of peanut and grain bar preloads on postmeal satiety, glycemia, and weight loss in health...