cDNA cloning of porcine brain prolyl endopeptidase and identification of the active-site seryl residue

Journal Article · Tue Feb 26 04:00:00 EST 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00222a025· OSTI ID:5745430

Rennex, D; Hemmings, B A; Hofsteenge, J; Stone, S R ^[1]

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1. Friedrich Miescher-Institut, Basel (Switzerland)

Prolyl endopeptidase is a cytoplasmic serine protease. The enzyme was purified from porcine kidney, and oligonucleotides based on peptide sequences from this protein were used to isolate a cDNA clone from a porcine brain library. This clone contained the complete coding sequence of prolyl endopeptidase and encoded a polypeptide with a molecular mass of 80751 Da. The deduced amino acid sequence of prolyl endopeptidase showed no sequence homology with other known serine proteases. ({sup 3}H)Diisopropyl fluorophosphate was used to identify the active-site serine of prolyl endopeptidase. One labeled peptide was isolated and sequenced. The sequence surrounding the active-site serine was Asn-Gly-Gly-Ser-Asn-Gly-Gly. This sequence is different from the active-site sequences of other known serine proteases. This difference and the lack of overall homology with the known families of serine proteases suggest that prolyl endopeptidase represents a new type of serine protease.

• OSTI ID: 5745430
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 30:8; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
ANIMALS
BODY
BRAIN
CENTRAL NERVOUS SYSTEM
CLONING
DNA
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
DOMESTIC ANIMALS
ELECTROPHORESIS
ENZYME ACTIVITY
ENZYMES
HYBRIDIZATION
HYDROGEN COMPOUNDS
HYDROLASES
MAMMALS
MOLECULAR STRUCTURE
NERVOUS SYSTEM
NUCLEIC ACIDS
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
ORGANS
PEPTIDE HYDROLASES
PROTEINS
PURIFICATION
RECOMBINANT DNA
SERINE PROTEINASES
STRUCTURAL CHEMICAL ANALYSIS
SWINE
TRITIUM COMPOUNDS
VERTEBRATES