Two monoclonal antibodies (MAbs) to a 36-kDa extracellular metalloprotease (PSCP) from Burkholderia (Pseudomonas) cepacia were found to react with thermolysin, Pseudomonas aeruginosa elastase, alkaline protease (Apr) and LasA, Serratia marcescens protease (SMP), Aeromonas hydrophila protease (AhP), and both the lethal factor (LF) and protective antigen (PA) of Bacillus anthracis on immunoblots. The MAbs were capable of neutralising the proteolytic activity of thermolysin, P. aeruginosa elastase and PSCP but not that of Apr, SMP, and AhP. These results suggest that these MAbs may be able to differentiate between the thermolysin and serralysin family of metalloproteases on the basis of their neutralisation capability and could, therefore, be useful tools in the characterisation of new bacterial proteases.
MoffatJ. F.,
EdelsteinP. H.,
RegulaD. P.,
CirilloJ. D.,
TompkinsL. S. Effects of an isogenic Zn-metalloprotease-deficient mutant of Legionella pneumophila in a guinea-pig pneumonia model. Mol Microbiol1994; 12:693–705
FinkelsteinR. A.,
Boesman-FinkelsteinM.,
HoltP.Vibrio cholerae hemagglutinin/lectin/protease hydrolyzes fibronectin and ovomucin: F. M. Burnet revisited. Proc Natl Acad Sci USA1983; 80:1092–1095
BeverR. A.,
IglewskiB. H. Molecular characterization and nucleotide sequence of the Pseudomonas aeruginosa elastase structural gene. J Bacteriol1988; 170:4309–4314
DuongF.,
LazdunskiA.,
CamiB.,
MurgierM. Sequence of a cluster of genes controlling synthesis and secretion of alkaline protease in Pseudomonas aeruginosa; relationships to other secretory pathways. Gene1992; 121:47–54
BritiganB. E.,
HayekM. B.,
DoebbelingB. N.,
FickR. B. Transferrin and lactoferrin undergo proteolytic cleavage in Pseudomonas aeruginosa-infected lungs of patients with cystic fibrosis. Infect Immun1993; 61:5049–5055
HäseC. C.,
FinkelsteinR. A. Cloning and nucleotide sequence of the Vibrio cholerae hemagglutinin/protease (HA/protease) gene and construction of an HA/protease-negative strain. J Bacteriol1991; 173:3311–3317
MoffatJ. F.,
BlackW. J.,
TompkinsL. S. Further molecular characterization of the cloned Legionella pneumophila zinc metalloprotease. Infect Immun1994; 62:751–753
BodeW.,
Gomis-Rüth, F-X., StöckerW. Astacins, serralysins, snake venom and matrix metalloproteases exhibit identical zinc-binding environments (HEXXHXXGXXH and met-tum) and topologies and should be grouped into a common family, the ‘metaincins’. FEBS letters1993; 331:134–140
LylerlyD.,
GrayL.,
KregerA. Characterization of rabbit corneal damage produced by serratia keratitis and by a serratia protease. Infect Immun1981; 33:927–932
LylerlyD. M.,
KregerA. S. Importance of serratia protease in the pathogenesis of experimental Serratia marcescens pneumonia. Infect Immun1983; 40:113–119
OlsonJ. C.,
OhmanD. E. Efficient production and processing of elastase and LasA by Pseudomonas aeruginosa require zinc and calcium ions. J Bacteriol1992; 174:4140–4147
LoewyA. G.,
SanterU. V.,
WieczorekM.,
BlodgettJ. K.,
JonesS. W.,
CheronisJ. C. Purification and characterization of a novel zinc-proteinase from cultures of Aeromonas hydrophila. J Biol Chem1993; 268:9071–9078
SchiavoG.,
PoulainB.,
RossettoO.,
BenfenatiF.,
TaucL.,
MontecuccoC. Tetanus toxin is a zinc protein and its inhibition of neurotransmitter release and protease activity depend on zinc. EMBO J1992; 11:3577–3583
SchiavoG.,
ShoneC. C.,
RossettoO.,
AlexanderF. C. G.,
MontecuccoC. Botulinum neurotoxin serotype F is a zinc endopeptidase specific for VAMP/synaptobrevin. J Biol Chem1993; 268:11516–11519
KlimpelK. R.,
AuroraN.,
LepplaS. H. Anthrax toxin lethal factor contains a zinc metalloprotease consensus sequence which is required for lethal toxin activity. Mol Microbiol1994; 13:1093–1100
McKevittA. I.,
BajaksouzianS.,
KlingerJ. D.,
WoodsD. E. Purification and characterization of an extracellular protease from Pseudomonas cepacia. Infect Immun1989; 57:771–778
TowbinH.,
StaehelinT.,
GordonJ. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA1979; 76:4350–4354
RinderknechtH.,
GeokasM. C.,
SilvermanP.,
HaverbackB. J. A new ultra-sensitive method for determination of proteolytic activity. Clin Chim Acta1968; 21:197–203