Identification and characterization of the metal ion-dependent L-alanoyl-D-glutamate peptidase encoded by bacteriophage T5

The FEBS Journal
Galina V MikoulinskaiaOlga A Stepnaya

Abstract

Although bacteriophage T5 is known to have lytic proteins for cell wall hydrolysis and phage progeny escape, their activities are still unknown. This is the first report on the cloning, expression and biochemical characterization of a bacteriophage T5 lytic hydrolase. The endolysin-encoding lys gene of virulent coliphage T5 was cloned in Escherichia coli cells, and an electrophoretically homogeneous product of this gene was obtained with a high yield (78% of total activity). The protein purified was shown to be an L-alanoyl-D-glutamate peptidase. The enzyme demonstrated maximal activity in diluted buffers (25-50 mM) at pH 8.5. The enzyme was strongly inhibited by EDTA and BAPTA, and fully reactivated by calcium/manganese chlorides. It was found that, along with E. coli peptidoglycan, peptidase of bacteriophage T5 can lyse peptidoglycans of other Gram-negative microorganisms (Pectobacterium carotovorum, Pseudomonas putida, Proteus vulgaris, and Proteus mirabilis). This endolysin is the first example of an L-alanoyl-D-glutamate peptidase in a virulent phage infecting Gram-negative bacteria. There are, however, a great many sequences in databases that are highly similar to that of bacteriophage T5 hydrolase, indicating a wide dist...Continue Reading

References

Sep 4, 1992·Biochimica Et Biophysica Acta·J Caldentey, D H Bamford
Dec 12, 1989·Biochemistry·K S Larsen, D S Auld
Jan 1, 1988·Methods in Enzymology·D S Auld
Jul 25, 1986·Nucleic Acids Research·D S McPheetersL Gold
Jun 1, 1973·The Biochemical Journal·J B Ward
Nov 11, 1983·Nucleic Acids Research·F BrunelJ Davison
Jan 1, 1995·Methods in Enzymology·N D Rawlings, A J Barrett
Jan 1, 1995·Methods in Enzymology·D S Auld
Jan 1, 1995·FEMS Microbiology Letters·J C QuintelaM A de Pedro
Feb 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·J Wang, M D Cooper
Dec 15, 1995·The Biochemical Journal·A AballayL S Mayorga
Sep 1, 1997·Nucleic Acids Research·S F AltschulD J Lipman
Nov 12, 1998·Trends in Biochemical Sciences·F JeanmouginT J Gibson
May 25, 1999·FEBS Letters·K DüringW Gieffers
May 2, 2000·Applied and Environmental Microbiology·I AhrenholtzW Wackernagel
Mar 22, 2001·Proceedings of the National Academy of Sciences of the United States of America·D NelsonV A Fischetti
Aug 23, 2002·Nature·Raymond SchuchVincent A Fischetti
Nov 20, 2002·Research in Microbiology·Thomas G BernhardtRy Young
Jan 9, 2004·Applied and Environmental Microbiology·Stéphanie-Marie DeutschSylvie Lortal
Apr 20, 2004·Proceedings of the National Academy of Sciences of the United States of America·Min XuRy Young
Jul 17, 2004·Microbiology·David G PritchardJeffrey A Engler
Jan 22, 2005·Virology·Jianbin WangSongnian Hu
Jun 28, 2005·Current Opinion in Microbiology·Martin J Loessner
Mar 28, 2006·Experimental Biology and Medicine·Jan BorysowskiAndrzej Górski
May 10, 2007·Molecular Biology and Evolution·Koichiro TamuraSudhir Kumar
Jun 26, 2007·Molecular Genetics and Genomics : MGG·Tatsuya FukushimaJunichi Sekiguchi
Sep 29, 2007·Journal of Molecular Biology·Elizabeth J SummerRy Young
Oct 13, 2007·The Journal of Biological Chemistry·Yoshikuni GotoMasafumi Tsujimoto

❮ Previous
Next ❯

Citations

Feb 15, 2011·Extremophiles : Life Under Extreme Conditions·Mengxian LongXun Xu
Feb 15, 2013·Journal of Virology·Hugo OliveiraLeon D Kluskens
Jan 12, 2013·Current Protein & Peptide Science·Zuzanna Drulis-KawaRob Lavigne
Feb 1, 2012·Research in Microbiology·Jeong-A LimSangryeol Ryu
Apr 26, 2016·Journal of Biomolecular Structure & Dynamics·Victor P KutyshenkoVladimir N Uversky
Jul 5, 2016·Biochimica Et Biophysica Acta·Victor P KutyshenkoVladimir N Uversky
Mar 7, 2018·Journal of Molecular Microbiology and Biotechnology·Carlos Eduardo Serrano-MaldonadoMaricarmen Quirasco
Mar 15, 2019·Metallomics : Integrated Biometal Science·Angelina O KovalenkoGalina V Mikoulinskaia
Nov 13, 2015·Standards in Genomic Sciences·Yongle XuNianzhi Jiao
Jun 24, 2017·Annual Review of Virology·Kimi AzadJohn E Johnson
Sep 5, 2020·International Journal of Molecular Sciences·Jaclyn G McCutcheonJonathan J Dennis
Oct 22, 2016·Applied Microbiology and Biotechnology·Barbara MaciejewskaZuzanna Drulis-Kawa
Oct 28, 2020·Journal of Applied Microbiology·V S ShadrinG V Mikoulinskaia
Nov 8, 2020·International Journal of Biological Macromolecules·Victor P KutyshenkoVladimir N Uversky

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.

Bacterial Cell Wall Structure (ASM)

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.