Biomaterial Database

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. 1989

P M Fischer, and M E Howden

Division of Biological and Health Sciences, Deakin University, Geelong, Victoria, Australia.

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.

UI	MeSH Term	Description	Entries
D008832	Microchemistry	The development and use of techniques and equipment to study or perform chemical reactions, with small quantities of materials, frequently less than a milligram or a milliliter.
D002851	Chromatography, High Pressure Liquid	Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.	Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D004124	p-Dimethylaminoazobenzene	A reagent used mainly to induce experimental liver cancer. According to the Fourth Annual Report on Carcinogens (NTP 85-002, p. 89) published in 1985, this compound "may reasonably be anticipated to be a carcinogen." (Merck, 11th ed)	Butter Yellow,Dimethylaminoazobenzene,4-Dimethylaminoazobenzene,Methyl Yellow,p-Dimethylaminoazobenzene, (E)-Isomer,p-Dimethylaminoazobenzene, (Z)-Isomer,4 Dimethylaminoazobenzene,p Dimethylaminoazobenzene
D006827	Hydantoins	Compounds based on imidazolidine dione. Some derivatives are ANTICONVULSANTS.	Hydantoin,Imidazolidine-2,4-Diones,Imidazolidine 2,4 Diones
D000595	Amino Acid Sequence	The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.	Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000596	Amino Acids	Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.	Amino Acid,Acid, Amino,Acids, Amino
D013861	Thiocyanates	Organic derivatives of thiocyanic acid which contain the general formula R-SCN.	Rhodanate,Rhodanates
D013867	Thiohydantoins	Sulfur analogs of hydantoins with one or both carbonyl groups replaced by thiocarbonyl groups.	Thiohydantoin
D017879	Isothiocyanates	Organic compounds with the general formula R-NCS.