Crystal structure of a dodecameric FMN-dependent UbiX-like decarboxylase (Pad1) from Escherichia coli O157: H7

Bacteriophage endolysins degrade peptidoglycan and have been identified as antibacterial candidates to combat antimicrobial resistance. Considering the catalytic and structural diversity of endolysins, there is a paucity of structural data to inform how these enzymes work at the molecular level—key data that is needed to realize the potential of endolysin-based antibacterial agents. Here, we determine the atomic structure and define the enzymatic function of Escherichia coli O157:H7 phage FTEBc1 endolysin, LysT84. Bioinformatic analysis reveals that LysT84 is a modular endolysin, which is unusual for Gram-negative endolysins, comprising a peptidoglycan binding domain and an enzymatic domain. The crystal structure of LysT84 (2.99 Å) revealed a mostly α-helical protein with two domains connected by a linker region but packed together. LysT84 was determined to be a monomer in solution using analytical ultracentrifugation. Small-angle X-ray scattering data revealed that LysT84 is a flexible protein but does not have the expected bimodal P(r) function of a multidomain protein, suggesting that the domains of LysT84 pack closely creating a globular protein as seen in the crystal structure. Structural analysis reveals two key glutamate residues positioned on either side of the active site cavity; mutagenesis demonstrating these residues are critical for peptidoglycan degradation. Molecular dynamic simulations suggest that the enzymatically active domain is dynamic, allowing the appropriate positioning of these catalytic residues for hydrolysis of the β(1–4) bond. Overall, our study defines the structural basis for peptidoglycan degradation by LysT84 which supports rational engineering of related endolysins into effective antibacterial agents.

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Crystal structure of ureidoglycolate hydrolase (AllA) from Escherichia coli O157:H7

Proteins Structure Function and Bioinformatics ◽

10.1002/prot.20537 ◽

2005 ◽

Vol 61 (2) ◽

pp. 454-459 ◽

Cited By ~ 8

Author(s):

Stephane Raymond ◽

Ante Tocilj ◽

Eunice Ajamian ◽

Yunge Li ◽

Ming-Ni Hung ◽

...

Keyword(s):

Crystal Structure ◽

Escherichia Coli ◽

Escherichia Coli O157

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Crystal structure and molecular mechanism of an aspartate/glutamate racemase from Escherichia coli O157

FEBS Letters ◽

10.1002/1873-3468.12148 ◽

2016 ◽

Vol 590 (8) ◽

pp. 1262-1269 ◽

Cited By ~ 7

Author(s):

Xiuhua Liu ◽

Fei Gao ◽

Yinliang Ma ◽

Shuang Liu ◽

Yaqi Cui ◽

...

Keyword(s):

Crystal Structure ◽

Escherichia Coli ◽

Molecular Mechanism ◽

Escherichia Coli O157 ◽

Glutamate Racemase

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Crystal structure and biochemical features of dye-decolorizing peroxidase YfeX from Escherichia coli O157 Asp 143 and Arg 232 play divergent roles toward different substrates

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2017.01.081 ◽

2017 ◽

Vol 484 (1) ◽

pp. 40-44 ◽

Cited By ~ 6

Author(s):

Xiuhua Liu ◽

Zenglin Yuan ◽

Jiaxu Wang ◽

Yaqi Cui ◽

Shuang Liu ◽

...

Keyword(s):

Crystal Structure ◽

Escherichia Coli ◽

Escherichia Coli O157 ◽

Biochemical Features

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Bedeutung von Escherichia coli O157 beim Schlachtschaf in der Schweiz

Schweizer Archiv für Tierheilkunde ◽

10.1024/0036-7281.148.6.289 ◽

2006 ◽

Vol 148 (6) ◽

pp. 289-295 ◽

Cited By ~ 8

Author(s):

C. Zweifel ◽

M. Kaufmann ◽

J. Blanco ◽

R. Stephan

Keyword(s):

Escherichia Coli ◽

Escherichia Coli O157

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Molecular profiling and antimicrobial susceptibility of Escherichia coli O157:H7 isolated in Bulgaria

BULGARIAN JOURNAL OF VETERINARY MEDICINE ◽

10.15547/bjvm.2263 ◽

2020 ◽

Vol 23 (3) ◽

pp. 310-318

Author(s):

K. Koev ◽

T. Stoyanchev ◽

G. Zhelev ◽

P. Marutsov ◽

K. Gospodinova ◽

...

Keyword(s):

Escherichia Coli ◽

Escherichia Coli O157 ◽

Acid Sensitivity ◽

Lower Extent ◽

Phenotype Analysis ◽

Anal Swab ◽

Amoxicillin Clavulanic Acid ◽

Primary Identification ◽

Stx1 Gene ◽

Cattle Farms

The purpose of this study was to detect the presence of shiga-toxin producing Escherichia coli (STEC) in faeces of healthy dairy cattle and to determine the sensitivity of isolates to several antimicrobial drugs. A total of 1,104 anal swab samples originating from 28 cattle farms were examined. After the primary identification, 30 strains were found to belong to serogroup О157. By means of conventional multiplex PCR, isolates were screened for presence of resistance genes stx1, stx2 and eaeА. Twenty-nine strains possesses amplicons with a size corresponding to genes stx2 and eaeA, one had amplicons also for the stx1 gene and one lacked amplicons of all three genes. Twenty-eight strains demonstrated amplicons equivalent to gene H7. The results from phenotype analysis of resistance showed preserved sensitivity to ceftriaxone, ceftazidime, cefotaxime, cephalothin, streptomycin, gentamicin, tetracycline, enrofloxacin and combinations sulfamethoxazole/trimethoprim and amoxicillin/clavulanic acid. Sensitivity to ampicillin was relatively preserved, although at a lower extent.

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