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Title: Human mast cell tryptase: Multiple cDNAs and genes reveal a multigene serine protease family

Abstract

Three different cDNAs and a gene encoding human skin mast cell tryptase have been cloned and sequenced in their entirety. The deduced amino acid sequences reveal a 30-amino acid prepropeptide followed by a 245-amino acid catalytic domain. The C-terminal undecapeptide of the human preprosequence is identical in dog tryptase and appears to be part of a prosequence unique among serine proteases. The differences among the three human tryptase catalytic domains include the loss of a consensus N-glycosylation site in one cDNA, which may explain some of the heterogeneity in size and susceptibility to deglycosylation seen in tryptase preparations. All three tryptase cDNAs are distinct from a recently reported cDNA obtained from a human lung mast cell library. A skin tryptase cDNA was used to isolate a human tryptase gene, the exons of which match one of the skin-derived cDNAs. The organization of the {approx}1.8-kilobase-pair tryptase gene is unique and is not closely related to that of any other mast cell or leukocyte serine protease. The 5{prime} regulatory regions of the gene share features with those of other serine proteases, including mast cell chymase, but are unusual in being separated from the protein-coding sequence by an intron. High-stringency hybridization of amore » human genomic DNA blot with a fragment of the tryptase gene confirms the presence of multiple tryptase genes. These findings provide genetic evidence that human mast cell tryptases are the products of a multigene family.« less

Authors:
; ; ; ;  [1]
  1. (Univ. of California, San Francisco (USA))
Publication Date:
OSTI Identifier:
5322959
Resource Type:
Journal Article
Resource Relation:
Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States); Journal Volume: 87:10
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; RECOMBINANT DNA; DNA SEQUENCING; SERINE PROTEINASES; AMINO ACID SEQUENCE; GENES; LYMPHOCYTES; MAST CELLS; PHOSPHORUS 32; SKIN; ANIMAL CELLS; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY; BODY FLUIDS; CONNECTIVE TISSUE CELLS; DAYS LIVING RADIOISOTOPES; DNA; ENZYMES; HYDROLASES; ISOTOPES; LEUKOCYTES; LIGHT NUCLEI; MATERIALS; MOLECULAR STRUCTURE; NUCLEI; NUCLEIC ACIDS; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; ORGANS; PEPTIDE HYDROLASES; PHOSPHORUS ISOTOPES; RADIOISOTOPES; SOMATIC CELLS; STRUCTURAL CHEMICAL ANALYSIS; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Vanderslice, P., Ballinger, S.M., Tam, E.K., Goldstein, S.M., Craik, C.S., and Caughey, G.H.. Human mast cell tryptase: Multiple cDNAs and genes reveal a multigene serine protease family. United States: N. p., 1990. Web. doi:10.1073/pnas.87.10.3811.
Vanderslice, P., Ballinger, S.M., Tam, E.K., Goldstein, S.M., Craik, C.S., & Caughey, G.H.. Human mast cell tryptase: Multiple cDNAs and genes reveal a multigene serine protease family. United States. doi:10.1073/pnas.87.10.3811.
Vanderslice, P., Ballinger, S.M., Tam, E.K., Goldstein, S.M., Craik, C.S., and Caughey, G.H.. 1990. "Human mast cell tryptase: Multiple cDNAs and genes reveal a multigene serine protease family". United States. doi:10.1073/pnas.87.10.3811.
@article{osti_5322959,
title = {Human mast cell tryptase: Multiple cDNAs and genes reveal a multigene serine protease family},
author = {Vanderslice, P. and Ballinger, S.M., Tam, E.K. and Goldstein, S.M. and Craik, C.S. and Caughey, G.H.},
abstractNote = {Three different cDNAs and a gene encoding human skin mast cell tryptase have been cloned and sequenced in their entirety. The deduced amino acid sequences reveal a 30-amino acid prepropeptide followed by a 245-amino acid catalytic domain. The C-terminal undecapeptide of the human preprosequence is identical in dog tryptase and appears to be part of a prosequence unique among serine proteases. The differences among the three human tryptase catalytic domains include the loss of a consensus N-glycosylation site in one cDNA, which may explain some of the heterogeneity in size and susceptibility to deglycosylation seen in tryptase preparations. All three tryptase cDNAs are distinct from a recently reported cDNA obtained from a human lung mast cell library. A skin tryptase cDNA was used to isolate a human tryptase gene, the exons of which match one of the skin-derived cDNAs. The organization of the {approx}1.8-kilobase-pair tryptase gene is unique and is not closely related to that of any other mast cell or leukocyte serine protease. The 5{prime} regulatory regions of the gene share features with those of other serine proteases, including mast cell chymase, but are unusual in being separated from the protein-coding sequence by an intron. High-stringency hybridization of a human genomic DNA blot with a fragment of the tryptase gene confirms the presence of multiple tryptase genes. These findings provide genetic evidence that human mast cell tryptases are the products of a multigene family.},
doi = {10.1073/pnas.87.10.3811},
journal = {Proceedings of the National Academy of Sciences of the United States of America; (United States)},
number = ,
volume = 87:10,
place = {United States},
year = 1990,
month = 5
}
  • This submission in the Gene Antigen Register section of Immunogenetics journal describes research in which fluorescence in situ hybridization (FISH) was performed to map and determine independently the precise location of the human tryptase-2 (TRYP2) and human granzyme A (HFSP) genes. The results indicate that both genes map to chromosome 5q11-q12 and define a new locus for cytotoxic lymphocyte granule tryptases. 14 refs, 2 figs.
  • The cloning, expression, purification and crystallization of recombinant human kallikrein 7, directly synthesized in the active form in E. coli, is described. Diffraction data were collected to 2.8 Å resolution from native crystals. Human kallikreins are a group of serine proteases of high sequence homology whose genes are grouped as a single cluster at chromosome 19. Although the physiological roles of kallikreins are generally still unknown, members of the kallikrein family have been clearly implicated in pathological situations such as cancer and psoriasis. Human kallikrein 7 (hK7) has been shown to be involved in pathological keratinization, psoriasis and ovarian cancer.more » In order to gain insight into the molecular structure of this protein, hK7 was crystallized after recombinant production in its folded and active form using a periplasmic secretion vector in Escherichia coli. The crystals belonged to the rhombohedral space group H32 and diffracted to 2.8 Å. The phase problem was solved by molecular replacement using the mouse kallikrein-related protein neuropsin. Completion of the model and structure refinement are under way.« less
  • The chymotrypsin-like family of serine protease genes includes several members that are expressed exclusively in subsets of hematopoietic cells. For example, human neutrophil elastase and cathepsin G are expressed only in myelomonocytic precursors, and cytotoxic-T-cell serine proteases are found only in cytotoxic lymphocytes. The authors have used a cathepsin G cDNA probe to clone two cathepsin G-like genes (designated CGL-1 and CGL-2) from a human genomic library. They have determined that CGL-1 is identical to a previously identified gene (known as CCPI, CTLA I, or cytotoxic serine protease B) that is expressed only in activated cytotoxic T lymphocytes. They showmore » here that cathepsin G, CGL-1, and CGL-2 are linked on an {approx}50-kilobase locus found on human chromosome 14 at band q11.2. This gene cluster maps to the same chromosomal band as the {alpha} and {delta} T-cell receptor genes; this region is involved in most chromosomal translocations and inversions that are specifically associated with T-cell malignancies.« less
  • The authors have isolated and sequenced cDNAs for corticosteroid binding globulin (CBG) prepared from human liver and lung mRNAs. The results indicate that CBG mRNA is relatively abundant in the liver but is also present in the lung, testis, and kidney. The liver CBG cDNA contains an open reading frame for a 405-amino acid (M/sub r/ 45,149) polypeptide. This includes a predominantly hydrophobic, leader sequence of 22 residues that precedes the known NH/sub 2/-terminal sequence of human CBG. We, therefore, predict that the mature protein is composed of 383 amino acids and is a polypeptide of M/sub r/ 42,646. Amore » second, in-frame, 72-base-pair cistron of unknown significance exists between the TAA termination codon for CBG and a possible polyadenylylation signal (AATAAA) located 16 nucleotides before the polyadenylylation site. The deduced amino acid sequence of mature CBG contains two cysteine residues and consensus sequences for the attachment of six possible N-linked oligosaccharide chains. The sequences of the human lung and liver CBG cDNAs differ by only one nucleotide within the proposed leader sequence,and they attribute this to a point mutation. No sequence homology was found between CBG and other steroid binding proteins, but there is a remarkable similarity between the amino acid sequences of CBG and of ..cap alpha../sub 1/-antitrypsin, and this extends to other members of the serpin (serine protease inhibitor) superfamily« less
  • Human skin and lung mast cells and rodent peritoneal cells contain a carboxypeptidase in their secretory granules. The authors have screened human lung cDNA libraries with a mouse mast cell carboxypeptidase A (MC-CPA) cDNA probe to isolate a near-full-length cDNA that encodes human MC-CPA. The 5{prime} end of the human MC-CPA transcript was defined by direct mRNA sequencing and by isolation and partial sequencing of the human MC-CPA gene. Human MC-CPA is predicted to be translated as a 417 amino acid preproenzyme which includes a 15 amino acid signal peptide and a 94-amino acid activation peptide. The mature human MC-CPAmore » enzyme has a predicted size of 36.1 kDa, a net positive charge of 16 at neutral pH, and 86% amino acid sequence identity with mouse MC-CPA. DNA blot analyses showed that human MC-CPA mRNA is transcribed from a single locus in the human genome. Comparison of the human MC-CPA with mouse MC-CPA and with three rat pancreatic carboxypeptidases shows that these enzymes are encoded by distinct but homologous genes.« less