scholarly journals A novel matrix metalloprotease-like enzyme (karilysin) of the periodontal pathogen Tannerella forsythia ATCC 43037

2010 ◽  
Vol 391 (1) ◽  
Author(s):  
Abdulkarim Y. Karim ◽  
Magdalena Kulczycka ◽  
Tomasz Kantyka ◽  
Grzegorz Dubin ◽  
Abeer Jabaiah ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Proteolytic Activity ◽  
Calcium Ions ◽  
Peptide Bond ◽  
Periodontal Pathogen ◽  
Matrix Metalloprotease ◽  
Tannerella Forsythia ◽  
C Terminus ◽  
Autoproteolytic Cleavage ◽  
Thermal Stability Of

AbstractProteases ofTannerella forsythia, a pathogen associated with periodontal disease, are implicated as virulence factors. Here, we characterized a matrix metalloprotease (MMP)-like enzyme ofT. forsythiareferred 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.

scholarly journals Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37

2017 ◽  
Vol 398 (3) ◽  
pp. 395-409 ◽  
Author(s):  
Lahari Koneru ◽  
Miroslaw Ksiazek ◽  
Irena Waligorska ◽  
Anna Straczek ◽  
Magdalena Lukasik ◽  
...  
Keyword(s):  
Bactericidal Activity ◽  
Biochemical Characterization ◽  
Proteolytic Enzymes ◽  
Active Form ◽  
Peptide Bond ◽  
Binding Pocket ◽  
Periodontal Pathogen ◽  
Tannerella Forsythia ◽  
Complement Proteins ◽  
Multidomain Structure

Abstract Tannerella forsythia is a periodontal pathogen expressing six secretory proteolytic enzymes with a unique multidomain structure referred to as KLIKK proteases. Two of these proteases, karilysin and mirolysin, were previously shown to protect the bacterium against complement-mediated bactericidal activity. The latter metalloprotease, however, was not characterized at the protein level. Therefore, we purified recombinant mirolysin and subjected it to detailed biochemical characterization. Mirolysin was obtained as a 66 kDa zymogen, which autoproteolytically processed itself into a 31 kDa active form via truncations at both the N- and C-termini. Further autodegradation was prevented by calcium. Substrate specificity was determined by the S1′ subsite of the substrate-binding pocket, which shows strong preference for Arg and Lys at the carbonyl side of a scissile peptide bond (P1′ residue). The protease cleaved an array of host proteins, including human fibronectin, fibrinogen, complement proteins C3, C4, and C5, and the antimicrobial peptide, LL-37. Degradation of LL-37 abolished not only the bactericidal activity of the peptide, but also its ability to bind lipopolysaccharide (LPS), thus quenching the endotoxin proinflammatory activity. Taken together, these results indicate that, through cleavage of LL-37 and complement proteins, mirolysin might be involved in evasion of the host immune response.

scholarly journals Nucleotides bind to the C-terminus of ClC-5

2006 ◽  
Vol 398 (2) ◽  
pp. 289-294 ◽  
Author(s):  
Leigh Wellhauser ◽  
Hsin-Hen Kuo ◽  
Fiona L. L. Stratford ◽  
Mohabir Ramjeesingh ◽  
Ling-Jun Huan ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Secondary Structure ◽  
Chloride Ion ◽  
Helical Structure ◽  
Biophysical Properties ◽  
C Terminus ◽  
Cd Studies ◽  
Cbs Domains ◽  
Chloride Ion Channels ◽  
Thermal Stability Of

Mutations in ClC-5 (chloride channel 5), a member of the ClC family of chloride ion channels and antiporters, have been linked to Dent's disease, a renal disease associated with proteinuria. Several of the disease-causing mutations are premature stop mutations which lead to truncation of the C-terminus, pointing to the functional significance of this region. The C-terminus of ClC-5, like that of other eukaryotic ClC proteins, is cytoplasmic and contains a pair of CBS (cystathionine β-synthase) domains connected by an intervening sequence. The presence of CBS domains implies a regulatory role for nucleotide interaction based on studies of other unrelated proteins bearing these domains [Ignoul and Eggermont (2005) Am. J. Physiol. Cell Physiol. 289, C1369–C1378; Scott, Hawley, Green, Anis, Stewart, Scullion, Norman and Hardie (2004) J. Clin. Invest. 113, 274–284]. However, to date, there has been no direct biochemical or biophysical evidence to support nucleotide interaction with ClC-5. In the present study, we have expressed and purified milligram quantities of the isolated C-terminus of ClC-5 (CIC-5 Ct). CD studies show that the protein is compact, with predominantly α-helical structure. We determined, using radiolabelled ATP, that this nucleotide binds the folded protein with low affinity, in the millimolar range, and that this interaction can be competed with 1 μM AMP. CD studies show that binding of these nucleotides causes no significant change in secondary structure, consistent with a model wherein these nucleotides bind to a preformed site. However, both nucleotides induce an increase in thermal stability of ClC-5 Ct, supporting the suggestion that both nucleotides interact with and modify the biophysical properties of this protein.

The Physical Properties of Tripeptide Stereocomplex Nano-Formations

2020 ◽  
Vol 16 (10) ◽  
pp. 1495-1503
Author(s):  
Alaa F. Nahhas ◽  
Alrayan F. Nahhas ◽  
Thomas J. Webster
Keyword(s):  
Thermal Stability ◽  
Aqueous Solution ◽  
Wound Healing ◽  
Physical Properties ◽  
Medical Applications ◽  
Physiological Conditions ◽  
New Class ◽  
Left Handed ◽  
C Terminus ◽  
Thermal Stability Of

The fast-growing use of supramolecular hydrogelators as biomaterials for a variety of applications, ranging from wound healing to drug delivery to tissue engineering, has highlighted the importance of synthetic design over recent years. Here, we report a new class of nanosheet stereocomplexes in aqueous solution and at physiological conditions (i. e., pH7.4), which are formed by physically mixing right- and left-handed tripeptide supramolecular hydrogelators without any external stimulus. Such tripeptides were obtained by incorporating either α-aminoisobutyric acid (Aib, U) or alanine (Ala, A) at the C-terminus or middle position of known peptide hydrogelators containing naphthalene and two phenylalanine residues. For hydrogels of these peptides, our results show that their morphologies and physical properties changed upon mixing with the L- and D-forms of the peptides forming suspension stereocomplexes. These interactions reduced molecular mobility by forming new structures with new properties and, therefore, increased the thermal stability of the compound promising for numerous medical applications.

The Ordered Phase Formation in Ti-Al-Ga Alloys

1970 ◽  
Vol 28 ◽  
pp. 440-441
Author(s):  
Shiro Fujishiro ◽  
Harold L. Gegel
Keyword(s):  
Thermal Stability ◽  
High Temperature ◽  
Titanium Alloys ◽  
Growth Kinetics ◽  
Stoichiometric Composition ◽  
Good Potential ◽  
Alpha Titanium ◽  
Cold Rolled ◽  
Kinetics Of ◽  
Thermal Stability Of

Ordered-alpha titanium alloys having a DO19 type structure have good potential for high temperature (600°C) applications, due to the thermal stability of the ordered phase and the inherent resistance to recrystallization of these alloys. Five different Ti-Al-Ga alloys consisting of equal atomic percents of aluminum and gallium solute additions up to the stoichiometric composition, Ti3(Al, Ga), were used to study the growth kinetics of the ordered phase and the nature of its interface.The alloys were homogenized in the beta region in a vacuum of about 5×10-7 torr, furnace cooled; reheated in air to 50°C below the alpha transus for hot working. The alloys were subsequently acid cleaned, annealed in vacuo, and cold rolled to about. 050 inch prior to additional homogenization

Microstructure studies of tungsten silicide schottky contacts on Gaas

1987 ◽  
Vol 45 ◽  
pp. 328-329
Author(s):  
Yih-Cheng Shih ◽  
E. L. Wilkie
Keyword(s):  
Thermal Stability ◽  
Field Effect Transistors ◽  
Schottky Contact ◽  
Schottky Contacts ◽  
Excellent Thermal Stability ◽  
Barrier Heights ◽  
Gaas Substrates ◽  
Film Adhesion ◽  
Threshold Voltages ◽  
Thermal Stability Of

Tungsten silicides (WSix) have been successfully used as the gate materials in self-aligned GaAs metal-semiconductor-field- effect transistors (MESFET). Thermal stability of the WSix/GaAs Schottky contact is of major concern since the n+ implanted source/drain regions must be annealed at high temperatures (∼ 800°C). WSi0.6 was considered the best composition to achieve good device performance due to its low stress and excellent thermal stability of the WSix/GaAs interface. The film adhesion and the uniformity in barrier heights and ideality factors of the WSi0.6 films have been improved by depositing a thin layer of pure W as the first layer on GaAs prior to WSi0.6 deposition. Recently WSi0.1 has been used successfully as the gate material in 1x10 μm GaAs FET's on the GaAs substrates which were sputter-cleaned prior to deposition. These GaAs FET's exhibited uniform threshold voltages across a 51 mm wafer with good film adhesion after annealing at 800°C for 10 min.

Thermal stability of Al-Cu-Fe icosahedral alloys

1991 ◽  
Vol 1 (12) ◽  
pp. 1823-1836 ◽  
Author(s):  
M. Bessière ◽  
A. Quivy ◽  
S. Lefebvre ◽  
J. Devaud-Rzepski ◽  
Y. Calvayrac
Keyword(s):  
Thermal Stability ◽  
Thermal Stability Of

Practical criteria for the thermal stability of a unidimensional superconductor

1994 ◽  
Vol 4 (4) ◽  
pp. 653-657
Author(s):  
B. Bonzi ◽  
M. El Khomssi ◽  
H. Lanchon-Ducauquis
Keyword(s):  
Thermal Stability ◽  
Thermal Stability Of

Time and thermal stability of magnetic properties of amorphous Fe80TM3B17 alloys

1998 ◽  
Vol 08 (PR2) ◽  
pp. Pr2-63-Pr2-66 ◽  
Author(s):  
R. Varga ◽  
P. Vojtaník ◽  
A. Lovas
Keyword(s):  
Thermal Stability ◽  
Magnetic Properties ◽  
Thermal Stability Of
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