Document Detail


Micropreparative gel electrophoresis of low-molecular-weight peptides: purification of highly insoluble amyloid peptide fragments.
MedLine Citation:
PMID:  8660494     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have used the continuous-elution micropreparative gel electrophoresis device described by Baumann and Lauraeus (Anal. Biochem. 214, 142-148, 1993) to purify low-molecular-weight peptide fragments from in-gel digested standard proteins as well as highly in-soluble amyloid peptides. Alzheimer's amyloid beta-peptide, gelsolin-derived amyloid peptide of the Finnish type, and a novel amyloid of the British type were purified from either homogenized brain or kidney tissue material to a high degree of purity in a single run. Using the high resolving capacity of the Tris-Tricine-SDS buffer system of Schaegger and von Jagow (Anal. Biochem. 166, 368-379, 1978) we were able to isolate two synthetic peptides with M(r)4329 and 3284, differing only by 1045 in mass. The total peptide recovery, as determined by amino acid sequence analysis and scanning densitometry, ranged between 60 and 80%. In order to demonstrate the utility of this technique we subjected some of the purified peptides to direct N-terminal amino acid sequence analysis, mass spectrometry, microbore high-performance liquid chromatography, and immunochemical studies. Our results show that micropreparative gel electrophoresis is an effective tool for the isolation of not only larger polypeptides but also small peptide fragments in a form suitable for further biological use.
Authors:
M Baumann; A Golabek; M Lalowski; T Wisniewski
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Analytical biochemistry     Volume:  236     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1996 May 
Date Detail:
Created Date:  1997-02-12     Completed Date:  1997-02-12     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  191-8     Citation Subset:  IM    
Affiliation:
Department of Pathology, New York University Medical Center, New York 10016, USA. marc.baumann@helsinki.fi
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Amyloid / isolation & purification
Amyloid beta-Protein / isolation & purification*
Electrophoresis, Polyacrylamide Gel / methods*
Membrane Proteins
Microchemistry / methods
Molecular Sequence Data
Molecular Weight
Peptide Fragments / isolation & purification*
Peptides / isolation & purification*
Solubility
Grant Support
ID/Acronym/Agency:
AG-10953/AG/NIA NIH HHS; NS-30445-04/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Amyloid; 0/Amyloid beta-Protein; 0/ITM2B protein, human; 0/Membrane Proteins; 0/Peptide Fragments; 0/Peptides; 0/amyloid AGel protein, human

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