CHAP domain of tail-associated protein from phage 812 and its
lytic activity against Staphylococcus aureus

a 2013

CHAP domain of tail-associated protein from phage 812 and its lytic activity against Staphylococcus aureus

BENEŠÍK, Martin, Radka DOPITOVÁ, Lubomír JANDA, Jiří DOŠKAŘ, Marek MOŠA et. al.

Základní údaje

Originální název

CHAP domain of tail-associated protein from phage 812 and its lytic activity against Staphylococcus aureus

Autoři

BENEŠÍK, Martin, Radka DOPITOVÁ, Lubomír JANDA, Jiří DOŠKAŘ, Marek MOŠA a Roman PANTŮČEK

Vydání

Molecular Genetics of Bacteria and Phages, 2013

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

Genetika a molekulární biologie

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Organizační jednotka

Přírodovědecká fakulta

Změněno: 1. 8. 2015 18:43, prof. RNDr. Roman Pantůček, Ph.D.

Anotace

V originále

Staphylococcus aureus is a major causative agent of human and animal diseases. The increasing number of pathogenic strains resistant to antimicrobial drugs is a serious public health problem that can be solved by applications of phage therapy as a suitable alternative to antibiotics treatment. Currently the research focus on the phage encoded antimicrobial proteins applicable to combat bacterial infections, such as phage endolysins that digest the cell wall of bacteria. In this study we focused on tail hydrolase of polyvalent staphylococcal bacteriophage 812, a member of SPO1-like viruses from family Myoviridae. This enzyme plays a role in the beginning of phage lytic cycle. It disturbs bacteria cell wall for injection of phage DNA into bacterial cell. CHAP domain was predicted at C-terminus of tail-associated protein by bioinformatics tools Total length of tail protein is 808 aa and CHAP domain is represented by 130 aa. The gene sequence for the CHAP domain was cloned and expressed as a soluble protein in E. coli. Expressed protein with his-tag was purified using metal-chelate affinity chromatography. Activity of this enzyme was verified on zymogram and using turbidity assay. Results show, that CHAP domain of tail-associated protein can lyse peptidogycan of S. aureus in zymograms. CHAP domain is also active against live bacterial cells and the protein decreas optical density of bacterial culture. Tail hydrolase is not as active like endolysin of phage 812. This problem can be resolved by modification of active site or by connecting CHAP domain with another domain.

Návaznosti

EE2.3.20.0183, projekt VaV

Název: Centrum experimentální biomedicíny

NT12395, projekt VaV

Název: Molekulární průkaz a analýza invazivních kmenů small colony variants (SCV) a rezistentních kmenů S. aureus od pacientů s cystickou fibrózou

TA01010405, projekt VaV

Název: Výzkum stafylokokových bakteriofágových mutant s širokým spektrem hostitelů (Akronym: TAČR/IMUNA-1)

Investor: Technologická agentura ČR, Výzkum stafylokokových bakteriofágových mutant s širokým spektrem hostitelů

Citovat

BENEŠÍK, Martin, Radka DOPITOVÁ, Lubomír JANDA, Jiří DOŠKAŘ, Marek MOŠA a Roman PANTŮČEK. CHAP domain of tail-associated protein from phage 812 and its lytic activity against Staphylococcus aureus. In Molecular Genetics of Bacteria and Phages. 2013.

@proceedings{1125494,
   author = {Benešík, Martin and Dopitová, Radka and Janda, Lubomír and Doškař, Jiří and Moša, Marek and Pantůček, Roman},
   booktitle = {Molecular Genetics of Bacteria and Phages},
   language = {eng},
   title = {CHAP domain of tail-associated protein from phage 812 and its lytic activity against Staphylococcus aureus},
   url = {http://www.union.wisc.edu/phages/},
   year = {2013}
}

TY  - CONF
ID  - 1125494
AU  - Benešík, Martin - Dopitová, Radka - Janda, Lubomír - Doškař, Jiří - Moša, Marek - Pantůček, Roman
PY  - 2013
TI  - CHAP domain of tail-associated protein from phage 812 and its lytic activity against Staphylococcus aureus
UR  - http://www.union.wisc.edu/phages/
N2  - Staphylococcus aureus is a major causative agent of human and animal diseases. The increasing number of pathogenic strains resistant to antimicrobial drugs is a serious public health problem that can be solved by applications of phage therapy as a suitable alternative to antibiotics treatment. Currently the research focus on the phage encoded antimicrobial proteins applicable to combat bacterial infections, such as phage endolysins that digest the cell wall of bacteria. In this study we focused on tail hydrolase of polyvalent staphylococcal bacteriophage 812, a member of SPO1-like viruses from family Myoviridae. This enzyme plays a role in the beginning of phage lytic cycle. It disturbs bacteria cell wall for injection of phage DNA into bacterial cell. CHAP domain was predicted at C-terminus of tail-associated protein by bioinformatics tools Total length of tail protein is 808 aa and CHAP domain is represented by 130 aa. The gene sequence for the CHAP domain was cloned and expressed as a soluble protein in E. coli. Expressed protein with his-tag was purified using metal-chelate affinity chromatography. Activity of this enzyme was verified on zymogram and using turbidity assay. Results show, that CHAP domain of tail-associated protein can lyse peptidogycan of S. aureus in zymograms. CHAP domain is also active against live bacterial cells and the protein decreas optical density of bacterial culture. Tail hydrolase is not as active like endolysin of phage 812. This problem can be resolved by modification of active site or by connecting CHAP domain with another domain.
ER  -

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