Isolation and sequence of a rat chymotrypsin B gene

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

Resultado de la investigación: Article

99 Citas (Scopus)

Resumen

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.

Idioma originalEnglish
Páginas (desde-hasta)14265-14270
Número de páginas6
PublicaciónJournal of Biological Chemistry
Volumen259
N.º22
EstadoPublished - 1984

Huella dactilar

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

Citar esto

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. En: Journal of Biological Chemistry. 1984 ; Vol. 259, N.º 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, n.º 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.

En: Journal of Biological Chemistry, Vol. 259, N.º 22, 1984, p. 14265-14270.

Resultado de la investigación: Article

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AU - Bell, G. I.

AU - Quinto, C.

AU - Quiroga, M.

AU - Valenzuela, P.

AU - Craik, C. S.

AU - Rutter, W. J.

PY - 1984

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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.