| Analysis of 4-N,N-dimethylaminoazobenzene 4'-thiohydantoin amino acids at sub-picomole levels by high-performance liquid chromatography: simultaneous manual sequencing of picomole quantities of several polypeptides. | |
MedLine Citation:
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PMID: 2500874 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A single-column high-performance liquid chromatographic separation of 4-N,N-dimethylaminoazobenzene 4'-thiohydantoin amino acid derivatives, generated during polypeptide sequence analysis by the 4-N,N-dimethylaminoazobenzene 4'-isothiocyanate/phenylisothiocyanate double coupling technique, is described. Recovery of the serine and threonine derivatives was improved by substituting boron trifluoride-diethyl etherate for trifluoroacetic acid in the thiazolinone cleavage reactions. Residues, including the S-carboxymethyl derivative of cysteine, were assigned after a single injection and a cycle time of 30 min. Quantities of 4-N,N-dimethylaminoazobenzene 4'-thiohydantoin amino acid derivatives as low as 100 fmol were detected. Interference of sequencing artefacts with residue assignment was avoided. This technique allows simultaneous manual sequencing of several proteins or peptides at the level of a few picomoles. |
Authors:
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P M Fischer; M E Howden |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Analytical biochemistry Volume: 177 ISSN: 0003-2697 ISO Abbreviation: Anal. Biochem. Publication Date: 1989 Feb |
Date Detail:
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Created Date: 1989-08-10 Completed Date: 1989-08-10 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 46-9 Citation Subset: IM |
Affiliation:
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Division of Biological and Health Sciences, Deakin University, Geelong, Victoria, Australia. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence* Amino Acids / analysis* Chromatography, High Pressure Liquid Hydantoins / analysis* Isothiocyanates* Microchemistry Thiocyanates Thiohydantoins / analysis* p-Dimethylaminoazobenzene / analogs & derivatives*, analysis |
| Chemical | |
Reg. No./Substance:
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0/Amino Acids; 0/Hydantoins; 0/Isothiocyanates; 0/Thiocyanates; 0/Thiohydantoins; 103-72-0/phenylisothiocyanate; 60-11-7/p-Dimethylaminoazobenzene; 72683-57-9/DABTH; 7612-98-8/4-(N,N-dimethylaminoazobenzene)-4'-isothiocyanate |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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