A novel matrix metalloprotease-like enzyme (karilysin) of the periodontal pathogen Tannerella forsythia ATCC 43037.
Proteases of Tannerella forsythia, a pathogen associated with periodontal disease, are implicated as virulence factors. Here, we characterized a matrix metalloprotease (MMP)-like enzyme of T. forsythia referred to as karilysin. Full-length (without a signal peptide) recombinant karilysin (49.9 kDa) processed itself into the mature 18-kDa enzyme through sequential autoproteolytic cleavage at both N- and C-terminal profragments. The first cleavage at the Asn14-Tyr15 peptide bond generated the fully active enzyme (47.9 kDa) and subsequent truncations at the C-terminus did not affect proteolytic activity. Mutation of Tyr15 to Ala generated a prokarilysin variant that processed itself into the final 18-kDa form with greatly reduced kinetics. Inactive prokarilysin with the mutated catalytic Glu residue (E136A) was processed by active karilysin at the same sites as the active enzymes. Karilysin proteolytic activity and autoprocessing were inhibited by 1,10-phenanthroline and EDTA. Calcium ions were found to be important for both the activity and thermal stability of karilysin. Using CLiPS technology, the specificity of karilysin was found to be similar to that of MMPs with preference for Leu/Tyr/Met at P1' and Pro/Ala at P3. This specificity and the ability to degrade elastin, fibrinogen and fibronectin may contribute to the pathogenicity of periodontitis.
Karim
Abdulkarim Y
AY
Department of Microbiology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, Krakow 30-387, Poland.
Kulczycka
Magdalena
M
Kantyka
Tomasz
T
Dubin
Grzegorz
G
Jabaiah
Abeer
A
Daugherty
Patrick S
PS
Thogersen
Ida B
IB
Enghild
Jan J
JJ
Nguyen
Ky-Anh
KA
Potempa
Jan
J
D000595:Amino Acid Sequence; D041963:Bacteroidetes / Q000201:enzymology*; D001483:Base Sequence; D006801:Humans; D053509:Matrix Metalloproteinase 13 / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010518:Periodontitis / Q000382:microbiology; D016415:Sequence Alignment; D013379:Substrate Specificity
D061965:Matrix Metalloproteinase Inhibitors; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases