Amino-terminal sequence analysis of the Coccidioides immitis chitinase/immunodiffusion-complement fixation protein

Abstract

A chitinase isolated from Coccidioides immitis was subjected to amino-terminal protein sequence analysis. The resulting 18-amino-acid sequence was compared with the previously reported amino acid sequence of coccidioidal immunodiffusion-complement fixation (IDCF) antigen. From the homology of the two sequences, the results support the identification of the IDCF antigen with a chitinase.

Molecular cloning and characterization of the Coccidioides immitis complement fixation/chitinase antigen

“…Nucleic acid sequencing showed that the cloned cDNA contains 1,623 bp, containing a single open reading frame of 1,281 bp, which encodes a protein of 427 amino acids having a predicted size of 47 kDa. A 35-amino-acid region, beginning at Ser-18 and ending at Arg-52, shows 92% homology with the reported Nterminal sequence of the mature, secreted CF/chitinase protein (15,21,22). The first 17 amino acids present in the deduced sequence of the cloned cDNA were not detected on the mature CF/chitinase protein.…”

“…Sequence comparisons of CF/chitinase cDNA with mature CF/chitinase protein. The deduced amino acid sequence of the cloned CF/chitinase cDNA was aligned with the N-terminal amino acid sequence which Johnson et al (15) reported for the mature CF/chitinase. As shown in Fig.…”

“…The primer set consisted of a forward primer based upon the N-terminal sequence of the CF/chitinase protein that was published by Johnson et al (15) and a reverse primer based upon a conserved internal amino acid sequence of chitinases from other fungi. As shown in Fig.…”

Use of a recombinant Coccidioides immitis complement fixation antigen-chitinase in conventional serological assays

“…Briefly, the clone was isolated from the cDNA library by using a 32 P-labeled, PCR-generated, chitinase-specific amplimer probe. This probe was generated from C. immitis genomic DNA by using primers synthesized from the combined CF-chitinase N-terminus amino acid sequence (11) and from the presumed bacterial and fungal internal active-site consensus amino acid sequence (1,6,12,23). The pCTS 4-2A insert DNA sequence is deposited in GenBank under accession no.…”

History of medical mycology in the united states

“…Resnick et al reported in 1987 the isolation and characterization of two major proteolytic components of C. immitis, a potent serine elastase and a metalloproteinase (337), and hypothesized their association with virulence. Johnson et al in 1993 isolated a chitinase from C. immitis spherule-endospores which was identical to the CF antigen, based on its biochemical properties and serologic activity (201). Amino-terminal protein sequence analysis of the chitinase linked the IDCF antigen with chitinase activity.…”