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PMID: 2783486 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cloning, analysis, and expression of murine perforin 1 cDNA, a component of cytolytic T-cell granules with homology to complement component C9.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 1 ·1989-01-00 ·Pages 247-51

Lowrey DM, Aebischer T, Olsen K, Lichtenheld M, Rupp F, Hengartner H, Podack ER

Abstract

The nucleotide sequence coding for the cytotoxic T-lymphocyte (CTL) protein perforin 1 (P1) has been determined and the corresponding protein sequence has been derived. Murine CTL cDNA libraries contained in the vector lambda gt11 were screened by using a monospecific antiserum to purified P1. Three recombinant phages were isolated and their cDNA inserts were sequenced. The derived protein sequence contains 554 amino acids and displays, as expected, considerable homology with certain functional domains in the complement components C9, C8 alpha, C8 beta, and C7. The identity of P1 cDNA clones was verified by prokaryotic expression and the reactivities of antisera produced to the expressed proteins. In addition, antisera were produced to two synthetic peptides located in the center and C-terminal portions of P1. All antisera reacted with purified P1. In Northern blot analyses, P1 cDNA probes recognized a 2.9-kilobase mRNA only in CTL. Perforin mRNA was found in all cloned CTL and in all mixed lymphocyte reactions that gave rise to cytotoxic cells. Perforin mRNA was also detected in virus-specific CTL that had been generated in vivo and isolated from liver tissue of mice infected with lymphocytic choriomeningitis virus. The cell-specific expression of perforin is consistent with its postulated role in cytolysis.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cloning, Molecular Complement C9/genetics Cytoplasmic Granules/immunology DNA/genetics Genes Humans Membrane Glycoproteins Membrane Proteins/genetics Mice Molecular Sequence Data Perforin Pore Forming Cytotoxic Proteins RNA, Messenger/genetics Sequence Homology, Nucleic Acid T-Lymphocytes, Cytotoxic/immunology Transcription, Genetic
Chemicals
Complement C9 Membrane Glycoproteins Membrane Proteins Pore Forming Cytotoxic Proteins RNA, Messenger Perforin DNA
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Lowrey D M
Department of Microbiology and Immunology, University of Miami, School of Medicine, FL 33101.
Aebischer T
Olsen K
Lichtenheld M
Rupp F
Hengartner H
Podack E R
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.1.247
Published
1989-01-00
Pages
247-51
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC286441
Subset
IM
Grants
NIAID NIH HHS · AI21999 · United States
NCI NIH HHS · CA39201 · United States
Databases
GENBANK
J04148
External Links
PubMed source DOI: 10.1073/pnas.86.1.247 PMC Full Text View journal details
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