Crystallization and preliminary X‐ray crystallographic studies of UDP‐<i>N</i>‐acetylenolpyruvylglucosamine reductase

Benson, Timothy E.; Walsh, Christopher T.; Hogle, James M.

doi:10.1002/pro.5560030718

Cited by 14 publications

(14 citation statements)

References 6 publications

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“…The monoclinic crystals did not show the wide variation in estimates of unit-cell dimensions found for the orthorhombic form. Diffraction characteristics of examples we grew of the tetragonal crystal form accord well with those reported (Benson et al, 1994). …”

Section: Data Collection and Analysissupporting

confidence: 85%

“…This other form could be grown under the same crystallization conditions as the monoclinic form, and a crystal size of about 0.3 x 0.2 × 0.2 mm could be attained with macroseeding. This crystal form proved to be identical to the tetragonal form reported by Benson and colleagues (Benson, Walsh & Hogle, 1994) and we later discovered the tetragonal form in some droplets containing UNAGEP that produced orthorhombic crystals.…”

Section: Crystallization and Derivative Preparationsupporting

confidence: 84%

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Crystallization and preliminary crystallographic analysis ofE. coliuridine 5'-diphospho-N-acetylenolpyruvylglucosamine reductase in two new crystal forms

Ohringer

Chang²,

Sheriff³

et al. 1996

Acta Cryst D

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Uridine 5'-diphospho-N-acetylenolpyruvylglucosamine reductase (MurB), the second enzyme in the peptidoglycan synthetic pathway of Escherichia coli, has been crystallized in two previously unreported forms, one orthorhombic and the other monoclinic. MurB (molecular mass 38kDa) crystallizes in a range of conditions that utilize polyethylene glycol fractions as precipitants, and crystals can be grown with or without the enzyme's substrate, uridine 5'-diphospho-N-acetylenolpyruvylglucosamine. X-ray diffraction from crystals of the orthorhombic form extends to 2 ,~ resolution and shows the symmetry and systematic absences of space group P2j2~2j. These crystals show significant variations in cell dimensions at room temperature and at 100 K. A crystal used to collect a 2.0,A resolution data set at a synchrotron source showed cell dimensions at ca 100K of a = 51.0, b---79.3 and c ---87.1 ,~, indicating one molecule peroasymmetric unit. The monoclinic crystals scatter X-rays to 3.0 A resolution consistent with space group P2~, unit-cell dimensions (ca 100K) a-----50.7, b=92.4, c--85.5A, and /5= 104 ° , and two molecules per asymmetric unit. Mercury derivatives have been prepared with both orthorhombic and monoclinic forms, and efforts are underway to exploit these derivatives to determine the structure of this protein.

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Section: Data Collection and Analysissupporting

confidence: 85%

Section: Crystallization and Derivative Preparationsupporting

confidence: 84%

Crystallization and preliminary crystallographic analysis ofE. coliuridine 5'-diphospho-N-acetylenolpyruvylglucosamine reductase in two new crystal forms

Ohringer

Chang²,

Sheriff³

et al. 1996

Acta Cryst D

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“…EDC was obtained from Aldrich (Milwaukee, WI), and HOBt was from Novabiochem (San Diego, CA). E. coli MurA and MurB were cloned and purified as previously described (12)(13)(14). All other chemicals were of the highest commercial grade available.…”

Section: Methodsmentioning

confidence: 99%

A Fluorescence-Based Homogeneous Assay for Measuring Activity of UDP–3-O-(R-3-Hydroxymyristoyl)-N-acetylglucosamine Deacetylase

Wang¹,

Maniar²,

Jain³

et al. 2001

Analytical Biochemistry

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“…Coculture of LGG microcapsules with E.coli appeared to down-regulate murB gene expression and to completely abolish expression of the murF gene. Bacteria generally synthesize their own active form of Nacetylmuramic acid, UDP-N-acetylmuramic acid [23], and the MurB enzyme (UDP-N-acetylglucosamine pyruvate enol ether reductase) plays an important role in the biosynthesis of this substance [24]. The MurB enzyme converts UDP-N-acetylglucosamine pyruvate enol ether to UDP-N-acetylmuramic acid by reducing the double bond in the enol ether structure [25].…”

Section: Effects Of Lgg Microcapsules On the Growth And Metabolism Ofmentioning

confidence: 99%

Label-free quantitative proteomic analysis of the inhibition effect of Lactobacillus rhamnosus GG on Escherich coli biofilm formation in coculture

Song

Lou

et al. 2020

Preprint

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Background: Escherich coli (E.coli) is the principal pathogen that causes biofilm formation; the latter is associated with infectious diseases and antibiotic resistance. In our previous work, we demonstrated that probiotic microcapsules have superior biofilm inhibition capacity compared to probiotic sterile culture supernatant. Herein, the mechanism of the inhibition effects was investigated using label-free quantitative proteomics analysis. Results:The proteomic analysis characterized a total of 1655 proteins in E.coli K12MG1655 and 1431 proteins in Lactobacillus rhamnosus GG (LGG). Among them, after coculture treatment, there were 262 and differentially expressed proteins that were specific for E.coli and 291 for LGG. The differentially expressed proteins after coculture were related to cellular metabolism, the stress response, transcription, and the cell membrane. In addition, we identified five strain-specific genes in E.coli and LGG, respectively, which were consistent with the proteomics results. Conclusions:These findings indicate that LGG microcapsules may inhibit E.coli biofilm inhibition by disrupting metabolic processes, particularly in relation to energy metabolism and stimulus responses, both of which are critical for the growth of LGG. Together, these findings increase our understanding of the interactions between bacteria under coculture conditions. BackgroundBiofilms are complex bacterial community structures that can attach to a surface. They are connected to this surface via extracellular polymeric substances (EPS) which form a matrix composed primarily of polysaccharides, proteins, and DNA; this encapsulates the bacteria [1]. Biofilms not only cause economic losses, but also present a public health hazard. This is because the bacteria present within biofilms are much more resistant to antibiotics, disinfectants [2], and the host immune system effectors [3]. Therefore, it is critical to uncover effective non-toxic, or less toxic, antifungal agents with novel modes of action.A recent study suggested that probiotic supernatants have antipathogenic properties [4], which implies that probiotics may inhibit biofilm formation through cell-cell communication. However, there

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Crystallization and preliminary X‐ray crystallographic studies of UDP‐N‐acetylenolpyruvylglucosamine reductase

Cited by 14 publications

References 6 publications

Crystallization and preliminary crystallographic analysis ofE. coliuridine 5'-diphospho-N-acetylenolpyruvylglucosamine reductase in two new crystal forms

Crystallization and preliminary crystallographic analysis ofE. coliuridine 5'-diphospho-N-acetylenolpyruvylglucosamine reductase in two new crystal forms

A Fluorescence-Based Homogeneous Assay for Measuring Activity of UDP–3-O-(R-3-Hydroxymyristoyl)-N-acetylglucosamine Deacetylase

Label-free quantitative proteomic analysis of the inhibition effect of Lactobacillus rhamnosus GG on Escherich coli biofilm formation in coculture

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