cDNA cloning and expression of guinea pig neutrophil attractant protein-1 (NAP-1)

NAP-1 is highly conserved in guinea pig

Teizo Yoshimura, D. G. Johnson

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

cDNA for neutrophil attractant protein-1 (NAP-1, also known as IL-8) was cloned from Con A-stimulated guinea pig spleen cells with human NAP-1 cDNA as a probe. Guinea pig NAP-1 cDNA is composed of 1433 bp with an open reading frame which encodes for a 101-amino-acid protein. Guinea pig NAP-1 had 70% amino acid sequence similarity to human NAP-1, which was much higher than a similarity between human and guinea pig monocyte chemoattractant protein-1 (MCP-1) (56%). Nucleotide sequence similarity within the coding region was 75%. To confirm its biological activity in guinea pig, recombinant guinea pig NAP-1 was expressed in COS-7 cells then purified. N-terminal sequence analysis gave two different N-termini at position 23 (Met) or 24 (Val). The two proteins showed their peak activity for guinea pig neutrophils at the concentration of 1 μg/ml (10 -7 M). Despite its high similarity to human NAP-1, the responsiveness of human neutrophils to guinea pig NAP-1 was minimum. Recombinant guinea pig NAP-1 caused strong neutrophil infiltration after intradermal injection into guinea pig skin. Since guinea pig is classified as a rodent, it was of interest to know whether human NAP-1 cDNA hybridizes to genomic DNA of other rodents such as mouse or rat, in which a NAP-1 homologue has not been found. Under low stringency conditions, human NAP-1 cDNA hybridized to human, rabbit and guinea pig DNA, but not to mouse or rat DNA. Unlike NAP-1, human MCP-1 cDNA hybridized to genomic DNA of rabbit, guinea pig, mouse, and rat; MCP-1 cDNA have been cloned from these species. The apparent absence of a NAP-1 gene in mouse or rat makes this chemoattractant unique among the members of the protein family to which NAP- 1 and MCP-1 belong.

Original languageEnglish
Pages (from-to)6225-6236
Number of pages12
JournalJournal of Immunology
Volume151
Issue number11
Publication statusPublished - 1993
Externally publishedYes

Fingerprint

Organism Cloning
Guinea Pigs
Neutrophils
Complementary DNA
Proteins
Chemokine CCL2
DNA
Rodentia
Rabbits
Intradermal Injections
Neutrophil Infiltration
COS Cells
Chemotactic Factors
Interleukin-8
Open Reading Frames
Sequence Analysis
Amino Acid Sequence
Spleen

ASJC Scopus subject areas

  • Immunology

Cite this

cDNA cloning and expression of guinea pig neutrophil attractant protein-1 (NAP-1) : NAP-1 is highly conserved in guinea pig. / Yoshimura, Teizo; Johnson, D. G.

In: Journal of Immunology, Vol. 151, No. 11, 1993, p. 6225-6236.

Research output: Contribution to journalArticle

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