Preparation of Tyr-C-peptide from genetically altered human insulin presursor

Journal Article · Fri Dec 01 04:00:00 EST 1995 · Applied Biochemistry and Biotechnology

DOI:https://doi.org/10.1007/BF02786858· OSTI ID:254424

Sun, Huaiyu; Tang, Jianguo; Hu, Meihao ^[1]

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1. Peking Univ., Beijing (China)

C-peptide radiommunoassay (C-peptide RIA) is widely used in determination of pancreatic B-cell secretion activity. {sup 125}I labeled Tyr-C-peptide is indispensable in C-peptide RIA kit. Herein we discuss a way of obtaining recombinant Tyr-C-peptide. Arg32Tyr human proinsulin mutant (R32Y-proinsulin) gene was constructed by site-directed mutagenesis and overexpressed in Escherichia coli. Purified R32Y-proinsulin was converted to insulin and Tyr-C-peptide by trypsin and carboxypeptidase B codigestion. Tyr-C-peptide was isolated through reverse-phase HPLC (RP-HPLC) and identified by C-peptide RIA and amino acid analysis. 12 refs., 5 figs., 1 tab.

• Sponsoring Organization: USDOE
• OSTI ID: 254424
• Journal Information: Applied Biochemistry and Biotechnology, Journal Name: Applied Biochemistry and Biotechnology Journal Issue: 3 Vol. 55; ISSN 0273-2289; ISSN ABIBDL
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
AMINO ACIDS
DNA-CLONING
ESCHERICHIA COLI
GENES
INSULIN
IODINE 125
MUTAGENESIS
OLIGONUCLEOTIDES
PEPTIDES
RADIOIMMUNOASSAY
STRUCTURE-ACTIVITY RELATIONSHIPS