Seiko Instruments Inc

Seiko Seiki Kabushiki Kaisha

Toyoaki Uchida

Akira Tsugita

Keiji Takamoto

Tatsuaki Ataka

Toshihiko Sakuhara

Ikuo Nanno

Masaharu Kamo

Kazuo Satake

Seitaro Takahashi

Yasuhiro Nomoto

A process is disclosed for sequencing proteins or peptides from the C-terminal end. The process comprises the steps of reacting the peptide or protein with an alkyl acid anhydride to convert the carboxy-terminal thereof into oxazolone, liberating the C-terminal amino acid by reaction with acid and alcohol or with ester, and identifying the liberated amino acid or amino acid derivative.

Method for amino acid sequencing of protein or peptide from carboxy

A protein or peptide which has an amino-terminus serine or threonine which has an acetylated alpha-amino group is allowed to react with an acid, and then allowed to react with an isothiocyanate under acidic conditions to thereby obtain a thiocarbamyl compound. Then the compound is to be analyzed using Edman degradation. Analysis can be performed with fewer operation steps and without using enzymes.

Method for sequencing of protein or peptide

A protein or peptide is treated by a vapor containing an organic acid represented by the following general formula CF.sub.3 --(CF.sub.2)n--COOH (n is zero or more integer). The resulting reaction mixture is processed by a mass spectrometer to obtain a mass spectrum to measure a mass of respective chemical species contained in the reaction mixture. Alternatively, the reaction mixture is processed by an amino acid analyzer to determine an amino acid sequence of the protein or peptide from the carboxy-terminal. According to another method, the protein or peptide is treated by an anhydride of the organic acid represented by the following general formula CF.sub.3 --(CF.sub.2)n--COOH (n is zero or more integer). The resulting reaction mixture is processed by a mass spectrometer to obtain a mass spectrum to measure a mass of respective chemical species contained in the reaction mixture to determine an amino acid sequence of the protein or peptide from the carboxy-terminal.

Method of determining amino acid sequence of protein or peptide from

An reaction vessel utilizes a sample carrier composed of a magnetic core and a surface coating effective to support sample of protein or peptide. The sample carrier is floated magnetically and positioned within a reaction chamber. Edman reagent is applied to the sample to effect amino acid sequence analysis of protein or peptide from amino-terminal. By such construction, reaction efficiency is increased to produce sequentially thiazolinon amino-acid derivatives to thereby increase number of identified remaining amino acids, thereby enabling microanalysis of sample.

Reaction vessel

A reaction chamber utilizes a sample carrier composed of a magnetic core and a surface coating effective to support sample of protein or peptide. The sample carrier is floated magnetically by means of electromagnets positioned within a reaction vessel. Edman reagnet is applied to the sample to effect amino acid sequence analysis of protein or peptide from amino-terminal. By such construction, reaction efficiency of repeated production of thiazolinon amino-acid derivatives is increased so as to increase number of identified amino acids, thereby enabling microanalysis of sample.

Reaction chamber for amino-terminal sequence analysis of proteins or

A method for detecting amino acid derivatives in which 2-anilino-5-thiazolinone derivatives of amino acids are reacted with an amino compound of fluorescein or rhodamine derivative amino compounds to form phenylthiocarbamyl amino acids derivatives. The phenylithiocarbamyl amino acids derivatives are detected with a high level of sensitivity by a fluorescence spectrophotometer.

Growing community of inventors

Tokyo, Japan

Toyoaki Uchida