Preparation of tyrosine-O-(/sup 35/S)sulfated cholecystokinin octapeptide from a nonsulfated precursor peptide

Journal Article · Tue Apr 01 00:00:00 EST 1986 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(86)90514-2· OSTI ID:5532149

Nakahara, T; Waki, M; Uchimura, H; Hirano, M; Kim, J S; Matsumoto, T; Nakamura, K; Ishibashi, K; Hirano, H; Shiraishi, A

A rapid and simple one-pot method for O-sulfation of nonsulfated cholecystokinin octapeptide (CCK-8) was developed using sulfuric acid and dicyclohexylcarbodiimide (DCC) without protection of the amino acid side chains. The extent of sulfation was increased with increasing the amount of reactants, sulfuric acid, and DCC, and reached maximum (40%) with fourfold molar excess of sulfuric acid and 40-fold molar excess of DCC. The excess of nonsulfated peptide inhibited the sulfation. The sulfation product was purified by HPLC or TLC to give a pure sulfated substance which showed exactly the same behavior as that of an authentic O-sulfated CCK-8 on HPLC or TLC. The purified sulfated peptide was active in stimulating amylase secretion from rat pancreatic fragments, and amino acid analysis showed that the tyrosine residue in the peptide existed in O-sulfated form. Sulfation with (/sup 35/S)sulfuric acid-DCC produced a radioactive substance, from which O-(/sup 35/S)sulfated CCK-8 could be easily purified by two-dimensional TLC.

• Research Organization: Kyushu Univ., Fukuoka, Japan
• OSTI ID: 5532149
• Journal Information: Anal. Biochem.; (United States), Vol. 154:1
• Country of Publication: United States
• Language: English

Related Subjects

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEAR CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
POLYPEPTIDES
CHEMICAL PREPARATION
LABELLING
AMYLASE
ENZYME INDUCTION
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
PANCREAS
PRECURSOR
PURIFICATION
RATS
SULFIDATION
SULFUR 35
SULFURIC ACID
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
TYROSINE
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ENDOCRINE GLANDS
ENZYMES
EVEN-ODD NUCLEI
GENE REGULATION
GLANDS
GLYCOSYL HYDROLASES
HYDROGEN COMPOUNDS
HYDROLASES
HYDROXY ACIDS
INORGANIC ACIDS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
NUCLEI
O-GLYCOSYL HYDROLASES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PEPTIDES
PROTEINS
RADIOISOTOPES
RODENTS
SEPARATION PROCESSES
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
400702* - Radiochemistry & Nuclear Chemistry- Properties of Radioactive Materials
550201 - Biochemistry- Tracer Techniques