Document Detail


Internal intensity standards for heme protein UV resonance Raman studies: excitation profiles of cacodylic acid and sodium selenate.
MedLine Citation:
PMID:  1846749     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We examine the utility of SO4(2-), ClO4-, cacodylic acid, and SeO4(2-) as internal intensity standards for Raman spectral measurements of protein structure. We find that 0.1 M SO4(2-) and ClO4- perturb the protein tertiary structure of aquomethemoglobin (met-Hb) and its fluoride (met-HbF) and azide (met-HbN3) complexes. Changes occur for the tryptophan near-UV absorption bands, the iron spin state is altered, and the fluoride ligand affinity decreases. Concentrations of ClO4- and SO4(2-) as low as 0.1 M suppress the met-HbF quaternary R----T transition induced by the allosteric effector inositol hexaphosphate (IHP). In contrast, similar concentrations of cacodylic acid and SeO4(2-) show little effect on the hemoglobin tertiary or quaternary protein structures or upon the R----T transition induced by IHP. We measure the Raman cross sections of cacodylic acid and SeO4(2-) between 218 and 514.5 nm and find that for UV excitation they are ca. 5-fold larger than ClO4- or SO4(2-). Thus, cacodylic acid and selenate can be used at lower concentrations. Cacodylic acid and SeO4(2-) are superior Raman internal intensity standards for protein structural studies.
Authors:
S H Song; S A Asher
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biochemistry     Volume:  30     ISSN:  0006-2960     ISO Abbreviation:  Biochemistry     Publication Date:  1991 Feb 
Date Detail:
Created Date:  1991-03-14     Completed Date:  1991-03-14     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1199-205     Citation Subset:  IM    
Affiliation:
Department of Chemistry, University of Pittsburgh, Pennsylvania 15260.
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MeSH Terms
Descriptor/Qualifier:
Cacodylic Acid / chemistry
Humans
Iron / chemistry
Methemoglobin / chemistry*
Perchloric Acid
Selenium / chemistry
Selenium Compounds*
Spectrophotometry, Ultraviolet
Spectrum Analysis, Raman / methods*
Sulfates / chemistry
Grant Support
ID/Acronym/Agency:
IR01GM30741-09/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Selenium Compounds; 0/Sulfates; 13410-01-0/sodium selenate; 7439-89-6/Iron; 75-60-5/Cacodylic Acid; 7601-90-3/Perchloric Acid; 7782-49-2/Selenium; 9008-37-1/Methemoglobin

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


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