Isolation and sequence of a rat chymotrypsin B gene

G. I. Bell, C. Quinto, M. Quiroga, P. Valenzuela, C. S. Craik, W. J. Rutter

Research output: Contribution to journalArticle

99 Citations (Scopus)

Abstract

A cDNA clone encoding part of chymotrypsin B was isolated from a cDNA library prepared from rat pancreatic mRNA and used as a probe to isolate the chymotrypsin B gene. The nucleotide sequence of this gene is presented. The 4709-base pair transcribed portion of the isolated gene was inferred from the cDNA and gene sequence, and the 5' border was determined by primer extension on pancreatic polyadenylated RNA. The coding portion of the gene is interrupted by six introns. The active site residues His 57, Asp 102, and Ser 195 are encoded by separate exons. Moreover, two regions of the enzyme which form the substrate-binding pocket are also encoded by separate exons. Thus, the substrate specificity and catalytic activity of the enzyme are produced by joining several exons encoding protein segments that are intrinsically catalytically inactive. The number and location of the intron/exon junctions of the chymotrypsin gene as compared to those of other serine protease genes, as well as the location of the genes on separate chromosomes, suggest that the duplication that resulted in the formation of the chymotrypsin gene was an ancient evolutionary event.

Original languageEnglish
Pages (from-to)14265-14270
Number of pages6
JournalJournal of Biological Chemistry
Volume259
Issue number22
Publication statusPublished - 1984

Fingerprint

Rats
Genes
Exons
Chymotrypsin
Introns
Complementary DNA
Chromosome Duplication
chymotrypsin B
Messenger RNA
Serine Proteases
Substrates
Enzymes
Chromosomes
Substrate Specificity
Gene Library
Base Pairing
Joining
Catalyst activity
Catalytic Domain
Nucleotides

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Bell, G. I., Quinto, C., Quiroga, M., Valenzuela, P., Craik, C. S., & Rutter, W. J. (1984). Isolation and sequence of a rat chymotrypsin B gene. Journal of Biological Chemistry, 259(22), 14265-14270.
Bell, G. I. ; Quinto, C. ; Quiroga, M. ; Valenzuela, P. ; Craik, C. S. ; Rutter, W. J. / Isolation and sequence of a rat chymotrypsin B gene. In: Journal of Biological Chemistry. 1984 ; Vol. 259, No. 22. pp. 14265-14270.
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Bell, GI, Quinto, C, Quiroga, M, Valenzuela, P, Craik, CS & Rutter, WJ 1984, 'Isolation and sequence of a rat chymotrypsin B gene', Journal of Biological Chemistry, vol. 259, no. 22, pp. 14265-14270.

Isolation and sequence of a rat chymotrypsin B gene. / Bell, G. I.; Quinto, C.; Quiroga, M.; Valenzuela, P.; Craik, C. S.; Rutter, W. J.

In: Journal of Biological Chemistry, Vol. 259, No. 22, 1984, p. 14265-14270.

Research output: Contribution to journalArticle

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N2 - A cDNA clone encoding part of chymotrypsin B was isolated from a cDNA library prepared from rat pancreatic mRNA and used as a probe to isolate the chymotrypsin B gene. The nucleotide sequence of this gene is presented. The 4709-base pair transcribed portion of the isolated gene was inferred from the cDNA and gene sequence, and the 5' border was determined by primer extension on pancreatic polyadenylated RNA. The coding portion of the gene is interrupted by six introns. The active site residues His 57, Asp 102, and Ser 195 are encoded by separate exons. Moreover, two regions of the enzyme which form the substrate-binding pocket are also encoded by separate exons. Thus, the substrate specificity and catalytic activity of the enzyme are produced by joining several exons encoding protein segments that are intrinsically catalytically inactive. The number and location of the intron/exon junctions of the chymotrypsin gene as compared to those of other serine protease genes, as well as the location of the genes on separate chromosomes, suggest that the duplication that resulted in the formation of the chymotrypsin gene was an ancient evolutionary event.

AB - A cDNA clone encoding part of chymotrypsin B was isolated from a cDNA library prepared from rat pancreatic mRNA and used as a probe to isolate the chymotrypsin B gene. The nucleotide sequence of this gene is presented. The 4709-base pair transcribed portion of the isolated gene was inferred from the cDNA and gene sequence, and the 5' border was determined by primer extension on pancreatic polyadenylated RNA. The coding portion of the gene is interrupted by six introns. The active site residues His 57, Asp 102, and Ser 195 are encoded by separate exons. Moreover, two regions of the enzyme which form the substrate-binding pocket are also encoded by separate exons. Thus, the substrate specificity and catalytic activity of the enzyme are produced by joining several exons encoding protein segments that are intrinsically catalytically inactive. The number and location of the intron/exon junctions of the chymotrypsin gene as compared to those of other serine protease genes, as well as the location of the genes on separate chromosomes, suggest that the duplication that resulted in the formation of the chymotrypsin gene was an ancient evolutionary event.

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Bell GI, Quinto C, Quiroga M, Valenzuela P, Craik CS, Rutter WJ. Isolation and sequence of a rat chymotrypsin B gene. Journal of Biological Chemistry. 1984;259(22):14265-14270.