A Protease Inhibitor of the Serpin Family Is a Major Protein in Carp Perimeningeal Fluid: II. cDNA Cloning, Sequence Analysis, and Escherichia coli Expression
Resource
Journal of Neurochemistry 64 (4): 1721-1727
Journal
Journal of Neurochemistry
Pages
1721-1727
Date Issued
1995
Date
1995
Author(s)
Abstract
Abstract: A cDNA clone, pCP9, has been isolated from a common carp liver cDNA library by immunoscreening with polyclonal antiserum raised against purified bighead carp α1‐antitrypsin. This clone is 1,396 bp in length and has an open reading frame encoding a protein of 410 amino acid residues. The deduced amino acid sequence shows moderate homology to human α1‐antitrypsin (38%), guinea pig contrapsin (35%), human α1‐antichymotrypsin (34%), and human proteinase C inhibitor (31%), all members of the serine protease inhibitor (serpin) family. To confirm further that the cDNA clone was derived from the authentic carp α1‐antitrypsin gene, the presumptive mature protein of pCP9 was expressed in Escherichia coli. The molecular mass of the recombinant protein matched that predicted from the nucleotide sequence. This recombinant protein, which was recognized by antiserum against native α1‐antitrypsin, was capable of formation of serpin‐enzyme complexes with trypsin, chymotrypsin, and elastase. Therefore, we conclude that the protein encoded by the pCP9 clone is indeed carp α1‐antitrypsin. Expression of α1‐antitrypsin in brain was confirmed by reverse transcription and polymerase chain reaction performed on mRNA derived from both common carp and bighead carp brain. Copyright © 1995, Wiley Blackwell. All rights reserved
Subjects
Carp; Perimeningeal fluid; Protease inhibitor; Serpin
Other Subjects
alpha 1 antichymotrypsin; alpha 1 antitrypsin; chymotrypsin; complementary dna; contrapsin; elastase; messenger rna; proteinase inhibitor; recombinant protein; serine proteinase inhibitor; trypsin; amino acid sequence; animal tissue; article; carp; cerebrospinal fluid; controlled study; dna library; dna sequence; enzyme inhibitor interaction; escherichia coli; guinea pig; human; molecular cloning; nonhuman; priority journal; protein determination; reverse transcription polymerase chain reaction; sequence homology; Amino Acid Sequence; Animal; Base Sequence; Body Fluids; Carps; Cloning, Molecular; DNA, Complementary; Escherichia coli; Meninges; Molecular Probes; Molecular Sequence Data; Nerve Tissue Proteins; Protease Inhibitors; Recombinant Proteins; RNA, Messenger; Sequence Homology; Serpins; Support, Non-U.S. Gov't
Type
journal article
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