Proteomic Analysis of the Sarcosine-Insoluble Outer Membrane Fraction of
            <i>Helicobacter pylori</i>
            Strain 26695

Baik, Seung‐Chul; Kim, Kyungmi; Song, Sumin; Kim, Do-Su; Jun, Jin-Su; Lee, Seung-Gyu; Song, Jae‐Young; Park, Jeong-Uck; Kang, Hyung-Lyun; Cho, Myung-Je; Youn, Hee‐Shang; Ko, Gyung‐Hyuck; Rhee, Kwang‐Ho

doi:10.1128/jb.187.4.1541.2005

Cited by 29 publications

(55 citation statements)

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“…The sample was focused with a maximum current of 50 μA, a maximum power of 200 mW and typical voltages ranging from 400 to 4000 V until 64 kVh was reached, after approximately 17 hrs (Baik et al, 2004;Ge et al, 2007). For the second dimension, samples from each of the 12 OFF-GEL fractions were separately mixed with sample buffer (4:1 ratio) containing 28.6% SDS and 4.76% 2-mercaptoethanol.…”

Section: Two-dimensional Gel Electrophoresismentioning

confidence: 99%

“…An in-gel digestion of the gel piece containing the protein band was performed using the method of O' Connell and Stalts (1997). First, the silver-stained excised gel piece was destained by adding 100 mM sodium thiosulphate and 30 mM potassium ferricyanide in a ratio of 1:1 for 20 min (Baik et al, 2004). Then, the supernatant was discarded and replaced by 100 μl of 200 mM ammonium bicarbonate for 20 min at room temperature.…”

Section: In-gel Digestion Of Proteins and Sample Clean Upmentioning

confidence: 99%

“…The resulting samples were cleaned up using ZipTipC18 pipette tips (Milipore). Finally, samples from the Zip-Tip were eluted into clean micro-centrifuge tubes and concentrated using a vacuum concentrator until they were completely dry (Eppendorf, USA), at this stage, the samples were ready for MALDI-TOF/ TOF mass analysis (Baik et al, 2004;Chan et al, 2006).…”

Section: In-gel Digestion Of Proteins and Sample Clean Upmentioning

confidence: 99%

See 2 more Smart Citations

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

Khalilpour

Agilan²,

Lee³

et al. 2013

Asian Pacific Journal of Cancer Prevention

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Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (≥ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culturepositive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/ TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline-5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.

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Section: Two-dimensional Gel Electrophoresismentioning

confidence: 99%

Section: In-gel Digestion Of Proteins and Sample Clean Upmentioning

confidence: 99%

Section: In-gel Digestion Of Proteins and Sample Clean Upmentioning

confidence: 99%

See 1 more Smart Citation

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

Khalilpour

Agilan²,

Lee³

et al. 2013

Asian Pacific Journal of Cancer Prevention

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“…Comparison of the provided data presents difficulties, due mainly to the genetic diversity of H. pylori as well as to methodological differences. There have been a number of significant attempts undertaken which were aimed at resolving surface or membrane proteins [5,8]. In each assay new immunoreactive proteins were identified and some of them have been tested as potential vaccine candidates.…”

Section: Introductionmentioning

confidence: 99%

Tip-α (hp0596 Gene Product) Is a Highly Immunogenic Helicobacter pylori Protein Involved in Colonization of Mouse Gastric Mucosa

et al. 2008

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A product of the Helicobacter pylori hp0596 gene (Tip-a) is a highly immunogenic homodimeric protein, unique for this bacterium. Cell fractionation experiments indicate that Tip-a is anchored to the inner membrane. In contrast, the three-dimensional model of the protein suggests that Tip-a is soluble or, at least, largely exposed to the solvent. hp0596 gene knockout resulted in a significant decrease in the level of H. pylori colonization as measured by real-time PCR assay. In addition, the Tip-a recombinant protein was determined to stimulate macrophage to produce IL-1a and TNF-a. Both results imply that Tip-a is rather loosely connected to the inner membrane and potentially released during infection.

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“…It is well known however, that H. pylori, which is a neutralophile, is able to survive by producing large amounts of the enzyme urease, which catalyzes hydrolysis of urea present in the stomach to yield NH 3 and CO 2 , thus elevating the pH to neutral as necessary for survival (6). It is generally accepted that urea hydrolysis is accomplished by uptake of urea through a proton-gated channel with hydrolysis taking place inside the bacterium and creating a thin neutral layer around the outer surface of the cell (7), although other studies indicate that this crucial enzyme exists on the cell surface (8)(9)(10), or that it is secreted or released (11,12). In addition to its ability to produce urease to elevate pH of its environment, H. pylori further protects itself by swimming through the protective layer of gastric mucus in the stomach and attaching to the epithelial cells beneath, where it can cause inflammation over the course of lifelong infection.…”

mentioning

confidence: 99%

Helicobacter pylori moves through mucus by reducing mucin viscoelasticity

Celli

Turner

Afdhal

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

385

326

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The ulcer-causing gastric pathogen Helicobacter pylori is the only bacterium known to colonize the harsh acidic environment of the human stomach. H. pylori survives in acidic conditions by producing urease, which catalyzes hydrolysis of urea to yield ammonia thus elevating the pH of its environment. However, the manner in which H. pylori is able to swim through the viscoelastic mucus gel that coats the stomach wall remains poorly understood. Previous rheology studies on gastric mucin, the key viscoelastic component of gastric mucus, indicate that the rheology of this material is pH dependent, transitioning from a viscous solution at neutral pH to a gel in acidic conditions. Bulk rheology measurements on porcine gastric mucin (PGM) show that pH elevation by H. pylori induces a dramatic decrease in viscoelastic moduli. Microscopy studies of the motility of H. pylori in gastric mucin at acidic and neutral pH in the absence of urea show that the bacteria swim freely at high pH, and are strongly constrained at low pH. By using two-photon fluorescence microscopy to image the bacterial motility in an initially low pH mucin gel with urea present we show that the gain of translational motility by bacteria is directly correlated with a rise in pH indicated by 2 ,7 -Bis-(2-Carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF), a pH sensitive fluorescent dye. This study indicates that the helicoidal-shaped H. pylori does not bore its way through the mucus gel like a screw through a cork as has previously been suggested, but instead achieves motility by altering the rheological properties of its environment.H. pylori ͉ bacterial motility ͉ rheology ͉ pH ͉ gelation

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Proteomic Analysis of the Sarcosine-Insoluble Outer Membrane Fraction of Helicobacter pylori Strain 26695

Cited by 29 publications

References 0 publications

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

Tip-α (hp0596 Gene Product) Is a Highly Immunogenic Helicobacter pylori Protein Involved in Colonization of Mouse Gastric Mucosa

Helicobacter pylori moves through mucus by reducing mucin viscoelasticity

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