PMID- 8112866
OWN - NLM
STAT- MEDLINE
DCOM- 19940330
LR  - 20210526
IS  - 0019-9567 (Print)
IS  - 1098-5522 (Electronic)
IS  - 0019-9567 (Linking)
VI  - 62
IP  - 3
DP  - 1994 Mar
TI  - Recombinant expression and antigenic properties of a 32-kilodalton extracellular 
      alkaline protease, representing a possible virulence factor from Aspergillus 
      fumigatus.
PG  - 936-42
AB  - A 32-kDa nonglycosylated alkaline protease (EC 3.4.1.14) with elastolytic 
      activity, secreted by the opportunistic pathogen Aspergillus fumigatus ATCC 
      42202, is suggested to be a virulence factor of this fungus. The enzyme is a 
      serine protease of the subtilisin family, and its cDNA nucleotide sequence has 
      recently been reported. We have cloned the cDNA encoding the mature protease into 
      a high-level Escherichia coli expression plasmid and produced the recombinant 
      protease as a fusion protein with a six-adjacent-histidine affinity tag at the 
      carboxy terminus. Subsequently, the recombinant protease was purified to 
      homogeneity, with affinity chromatography yielding 30 to 40 mg of recombinant 
      protease per liter of E. coli culture. Refolded recombinant protease, in 
      comparison with native protease, demonstrated weak enzymatic activity but similar 
      immunochemical characteristics as analyzed by antigen-specific enzyme-linked 
      immunosorbent assay (ELISA), competition ELISA, and immunoblotting assays. To 
      assess the allergenic potential of the protease, sera from patients with allergic 
      bronchopulmonary aspergillosis and sera from healthy control individuals were 
      analyzed by ELISA and immunoblotting techniques. Sera from patients with allergic 
      bronchopulmonary aspergillosis did not have protease-specific immunoglobulin E 
      (IgE) antibodies and, remarkably, did not show significantly elevated 
      protease-specific IgG antibody levels compared with those in sera from healthy 
      control individuals. This suggests that the alkaline protease from A. fumigatus 
      does not elicit IgE antibodies and has weak immunogenicity, a property which may 
      explain fungus persistence in allergic individuals.
FAU - Moser, M
AU  - Moser M
AD  - Swiss Institute of Allergy and Asthma Research, Davos-Platz.
FAU - Menz, G
AU  - Menz G
FAU - Blaser, K
AU  - Blaser K
FAU - Crameri, R
AU  - Crameri R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Infect Immun
JT  - Infection and immunity
JID - 0246127
RN  - 0 (Antigens, Bacterial)
RN  - 0 (Immunoglobulin G)
RN  - 0 (Recombinant Proteins)
RN  - EC 3.4.- (Endopeptidases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Antigens, Bacterial/*immunology
MH  - Aspergillus fumigatus/*enzymology/immunology/pathogenicity
MH  - Base Sequence
MH  - Endopeptidases/analysis/*biosynthesis/immunology
MH  - Humans
MH  - Immunoglobulin G/analysis
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Protein Folding
MH  - Recombinant Proteins/*biosynthesis
MH  - Virulence
PMC - PMC186206
EDAT- 1994/03/01 00:00
MHDA- 1994/03/01 00:01
PMCR- 1994/03/01
CRDT- 1994/03/01 00:00
PHST- 1994/03/01 00:00 [pubmed]
PHST- 1994/03/01 00:01 [medline]
PHST- 1994/03/01 00:00 [entrez]
PHST- 1994/03/01 00:00 [pmc-release]
AID - 10.1128/iai.62.3.936-942.1994 [doi]
PST - ppublish
SO  - Infect Immun. 1994 Mar;62(3):936-42. doi: 10.1128/iai.62.3.936-942.1994.