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J u l y 2 3 - 2 4 , 2 0 1 8 | R o m e , I t a l y
allied
academies
Joint Event on
Cardiology Congress 2018 & Microbe Infection 2018
Biomedical Research
|
ISSN: 0976-1683
|
Volume 29
2
nd
World Congress on
CARDIOLOGY
MICROBIOLOGY AND MICROBIAL INFECTION
&
39
th
Annual Congress on
Grzegorz Bereta et al., Biomed Res 2018, Volume 29 | DOI: 10.4066/biomedicalresearch-C1-003
GENETIC VARIABILITY OF PEPTIDYL
ARGININE DEIMINASE FROM
PORPHYROMONAS GINGIVALIS
IN
PERIODONTITIS PATIENTS
Grzegorz Bereta
1
, K Gawron
1
, Z Nowakowska
1
, K Łazarz-Barty-
zel
1
, M Chomyszyn-Gajewska
1
and
J Potempa
1,2
1
Jagiellonian University, Poland
2
University of Louisville School of Dentistry, USA
Introduction & Objective:
Periodontitis is a widespread chronic inflammatory
disease. Untreated condition leads to progressive destruction of the
periodontal tissue and may result in tooth loss. Changes in oral microbiome
leading to periodontitis are mainly driven by
Porphyromonas gingivalis
, the
pathogen producing numerous virulence factors, including peptidylarginine
deiminase (PPAD). PPAD modifies C-terminal arginine to citrulline, causing
changes in structure and function of modified proteins, contributing to
development periodontitis. The aim of this study was to investigate variability
of PPAD in clinical isolates of
P. gingivalis
.
Materials & Methods:
Together 23
P. gingivalis
strains were isolated from
patients with periodontitis and the PPAD gene was sequenced and analyzed
together with sequences extracted from the GenBank database. Identified
differences in the sequence were introduced into PPAD in reference strain
ATCC 33277 and expression (mRNA) and PPAD activity were measured in
cultures of the mutant. PPAD variants were expressed in
P. gingivalis
, purified
and used to compare their enzymatic properties. Clinical parameters of
periodontitis severity in patients infected with different
P. gingivalis
strains
were determined.
Results:
A new form of PPAD with three amino acid substitutions (G231N,
E232T, N235D) near the active site was found in approximately 30% of
P.
gingivalis
strains. Introduction of those mutations into the PPAD sequence
in the ATCC 33277 strain resulted in two-fold increase of PPAD activity in
culture, without effect on the level of mRNA expression. Kinetic assessment
of the enzymatic reaction revealed that the mutated form of PPAD had higher
maximum reaction rate (V
max
). Patients infected with
P. gingivalis
strains with
the super active PPAD variant had more advanced damage of periodontal
tissues.
Conclusion:
The newly identified form of PPAD shows higher enzymatic
activity and its presence in strains of
P. gingivalis
in periodontitis patients
correlated with severity of the disease.
Grzegorz Bereta has completed his MSc in Molecular
Biotechnology at Faculty of Biochemistry, Biophysics
and Biotechnology, Jagiellonian University, Krakow,
Poland and since then he is enrolled on PhD studies at
the same faculty. He has authored two publications and
two conference reports as well as one book chapter.
tnghd337@naver.comBIOGRAPHY