Document Detail


Characterization of neuropeptides extracted from canine intestine.
MedLine Citation:
PMID:  7688877     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Quantitative determination of neuropeptides in biologic tissues by radioimmunoassay requires both an efficient extraction of neuropeptides as well as maintenance of immunochemical reactivity. Vasoactive intestinal peptide, substance P, and met5-enkephalin were chosen for this study because they are neuropeptides which appear to be involved in multiple physiologic systems. Since all three neuropeptides have a methionine residue within their amino acid sequence, oxidation of methionine to methionine-sulfoxide during the extraction process could diminish their immunochemical reactivity. Multiple factors that might be important in extracting these neuropeptides from canine intestine, including pH of the solvent, tissue homogenization, heating, and addition of enzyme inhibitors, were examined. Concentrations of vasoactive intestinal peptide-like immunoreactivity and substance P-like immunoreactivity were significantly higher in acidic solvents, and tissue homogenization appeared to increase the concentrations of these two neuropeptides. Substance P-like immunoreactivity was increased by heating after tissue homogenization, suggesting heat-induced denaturation of tissue enzymes liberated by homogenization. Separation of acidic tissue extracts by high performance liquid chromatography followed by radioimmunoassay for all three neuropeptides revealed minor acid-induced oxidation of substance P. These results should be useful for planning the extraction of these three neuropeptides from other tissues.
Authors:
T R Koch; M F Otterson; D R Roddy; V L Go
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Neuropeptides     Volume:  24     ISSN:  0143-4179     ISO Abbreviation:  Neuropeptides     Publication Date:  1993 Jun 
Date Detail:
Created Date:  1993-09-10     Completed Date:  1993-09-10     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8103156     Medline TA:  Neuropeptides     Country:  SCOTLAND    
Other Details:
Languages:  eng     Pagination:  359-65     Citation Subset:  IM    
Affiliation:
Division of Gastroenterology, Zablocki VA Medical Center, Milwaukee, Wisconsin 53295.
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MeSH Terms
Descriptor/Qualifier:
Acetic Acid
Acetic Acids
Animals
Chromatography, High Pressure Liquid
Dogs
Enkephalin, Methionine / analysis,  isolation & purification*
Hot Temperature
Hydrochloric Acid
Hydrogen-Ion Concentration
Intestines / chemistry*
Oxidation-Reduction
Radioimmunoassay
Solvents
Substance P / analysis,  isolation & purification*
Vasoactive Intestinal Peptide / analysis,  isolation & purification*
Grant Support
ID/Acronym/Agency:
DK43104/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Acetic Acids; 0/Solvents; 33507-63-0/Substance P; 37221-79-7/Vasoactive Intestinal Peptide; 58569-55-4/Enkephalin, Methionine; 64-19-7/Acetic Acid; 7647-01-0/Hydrochloric Acid

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


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