Proteomic and Immunoblot Analyses of
            <i>Bartonella quintana</i>
            Total Membrane Proteins Identify Antigens Recognized by Sera from Infected Patients

Boonjakuakul, Jenni K.; Gerns, Helen L.; Chen, Yuting; Hicks, Linda D.; Minnick, Michael F.; Dixon, Scott E.; Hall, Steven C.; Koehler, Jane E.

doi:10.1128/iai.01974-06

Cited by 51 publications

(64 citation statements)

References 48 publications

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“…These proteins could be secreted by an unknown mechanism or released from bacteria with damaged cell membranes, an artifact of the purification step, and subsequently bound to the surface of intact cells. However, homologs of these proteins, including elongation factor G, polyribonucleotide nucleotidyltransferase, DnaK protein, chaperonin GroEL, two tRNA synthetases, aminopeptidase A, trigger factor, ATP synthase beta chain, dihydrolipoamide dehydrogenase, elongation factor Tu, recombinase A, and elongation factor Ts, were detected in the membrane fraction of R. conorii and other gramnegative bacteria (4,12,13,22,31,34). The localization of the WASP N-WASP MENA proteins or RickA on the cell surface of R. conorii has been demonstrated.…”

Section: Discussionmentioning

confidence: 99%

Proteomic Analysis of Rickettsia parkeri Strain Portsmouth

et al. 2009

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Rickettsia parkeri, a recently recognized pathogen of human, is one of several Rickettsia spp. in the United States that causes a spotted fever rickettsiosis. To gain insights into its biology and pathogenesis, we applied the proteomics approach to establish a two-dimensional gel proteome reference map and combined this technique with cell surface biotinylation to identify surface-exposed proteins of a low-passage isolate of R. parkeri obtained from a patient. We identified 91 proteins by matrix-assisted laser desorption ionization-tandem time of flight mass spectrometry. Of these, 28 were characterized as surface proteins, including virulence-related proteins (e.g., outer membrane protein A [OmpA], OmpB, ␤-peptide, and RickA). Two-dimensional immunoblotting with serum from the R. parkeri-infected index patient was utilized to identify the immunoreactive proteins as potential targets for diagnosis and vaccine development. In addition to the known rickettsial antigens, OmpA and OmpB, we identified translation initiation factor 2, cell division protein FtsZ, and cysteinyl-tRNA synthetase as immunoreactive proteins. The proteome map with corresponding cell surface protein analysis and antigen detection will facilitate a better understanding of the mechanisms of rickettsial pathogenesis.Rickettsia parkeri, a member of the spotted fever group Rickettsia (SFGR), was first isolated from the Gulf Coast tick, Amblyomma maculatum, in 1937 (29). In 2004, the first confirmed human infection with R. parkeri was reported in a 40-year-old man from the Tidewater area of coastal Virginia. The agent was isolated in cell culture from an eschar biopsy specimen and designated the Portsmouth strain (28). Recently, the first recognized case of tick bite-associated human infection was described (43); however, the epidemiology of R. parkeri is not well defined. In the United States, R. parkeri has been detected in A. maculatum and A. americanum; the geographical overlap between R. parkeri and these ticks with that of the vectors of R. rickettsii (the etiological agent of Rocky Mountain spotted fever [RMSF]) suggests that many cases of R. parkeri infection have been misidentified as RMSF (27,35). For example, Western blot analysis of serum specimens from 15 U.S. patients previously diagnosed with RMSF identified four serum specimens reactive with a 120-kDa protein of R. parkeri, suggesting infection with R. parkeri rather than R. rickettsii (30). However, a serologic test specific for this pathogen is not available (43), and little is known about its biology.Due to their obligate intracellular nature, genetic manipulation of Rickettsia has proven difficult. Alternatively, protein expression profiles (proteomes) are utilized to identify the mechanisms of pathogenesis and differentiate rickettsial species recognizing host immune response specificity to cell surface molecules, referred to as outer membrane proteins (Omps). The presence or absence of some Omps allows for differentiation between the typhus group and the SFGR, and the response...

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Section: Discussionmentioning

confidence: 99%

Proteomic Analysis of Rickettsia parkeri Strain Portsmouth

et al. 2009

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“…Todas as proteínas até então não têm sido descritas para outras riquétsias, sugerindo que representam proteínas imunologicamente reativas exclusivas de R. parkeri. No entanto, a antigenicidade de IF-2 e FtsZ tem sido relatado em outros trabalhos com diferentes espécies bacterianas de diferentes gêneros (Boonjakuakul et al 2007, Zhang et al 2008.…”

Section: Discussionunclassified

Detecção de proteínas imunorreativas de Rickettsia sp. cepa Mata Atlântica

et al. 2017

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RESUMO.-A Febre Maculosa Brasileira (FMB) é uma doença infecciosa, transmitida por carrapatos ao homem. Uma nova riquetsiose humana foi descrita como causadora de Febre Maculosa no Estado de São Paulo, sendo denominada de Rickettsia sp. cepa Mata Atlântica. O presente trabalho teve como objetivo detectar e identificar proteínas com potencial de estimular o sistema imune de hospedeiro mamí-fero, desta nova cepa descrita. Para tanto, foi realizado a extração proteica total de Rickettsia sp. cepa Mata Atlântica. As proteínas extraídas foram fracionadas por eletroforese. As bandas proteicas foram transferidas para membranas de nitrocelulose por migração elétrica e submetidas à téc-nica de Western-blot, para detecção proteica. Ao todo sete proteínas imunorreativas foram detectadas. Duas proteínas apresentaram maior abundancia, com peso molecular, de 200 e 130 kDa respectivamente. Através da comparação de mapas proteômicos existentes e pelo peso molecular que estas proteínas apresentaram, sugere-se que as duas proteínas detectadas representem rOmpA (200 kDa) e rOmpB (130 kDa). As demais proteínas detectadas apresentaram menor ocorrência e peso molecular inferior a 78 kDa, podendo representar membros da família de antígenos de superfície celular (Sca -Surface cell antigen). As proteínas detectadas poderão servir como base de estudo na elabo-

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“…Spots selected for protein identification were excised from GelCode Blue 1 stained gels followed by in-gel tryptic digestion and peptide extraction as previously published [Boonjakuakul et al, 2007]. Peptides were submitted to the Biomolecular Resource Center Mass Spectrometry Facility, UCSF, San Francisco, CA for protein identification.…”

Section: Preparation Of Peptidesmentioning

confidence: 99%

“…Protein identification was accomplished by using a combination of peptide mass fingerprinting (PMF) and high performance liquid chromatography (HPLC) electrospray ionization (ESI) tandem mass spectrometry (MS/MS) as described [Boonjakuakul et al, 2007]. For PMF protein identification, monoisotopic mass peak lists were submitted to the Aldente (http://expasy.org/tools/aldente/) search engine to facilitate protein identification.…”

Section: Mass Spectrometrymentioning

confidence: 99%

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Rapid alterations in cortical protein profiles underlie spontaneous sleep and wake bouts

Vázquez

Hall

Witkowska

et al. 2008

J of Cellular Biochemistry

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Existing data indicate that sleep-wakefulness is an essential behavior. The biological function(s) of sleep, however, remains unknown, due, in part, to the lack of information available at the intracellular level. Preliminary microarray analyses show that changes in behavioral state influence regional mRNA profiles; however, the impact of sleep on protein signatures is virtually unexplored. In these studies, cortical protein profiles were examined after timed bouts of spontaneous sleep-wakefulness. Within minutes of each behavioral state examined, a small number of spots showing unique expression were detected. Mass spectroscopy analyses of sleep-and wake-related spots identified proteins associated with multiple functional categories. Two sleep-associated proteins were further validated using a sleep deprivation paradigm. We found preliminary evidence for two different post-transcriptional mechanisms-one (GAPDH) in which the amount of protein was increased in the recovery sleep following prolonged waking, while the other (actin) suggested that post-translational modifications may underlie sleep. The similarities between the effects of sleep on both protein and mRNA profiles indicate that dynamic intracellular changes underlie sleep-wake states and are consistent with roles for sleep in multiple biological functions. J. Cell. Biochem. 105: 1472Biochem. 105: -1484Biochem. 105: , 2008. ß 2008 Wiley-Liss, Inc. KEY WORDS: TWO-DIMENSIONAL ELECTROPHORESIS (2DE); MASS SPECTROMETRY; SPONTANEOUS SLEEP-WAKE BOUTS; SLEEP-ASSOCIATED PROTEIN EXPRESSIONS leep-wake behavior is a complex and tightly regulated Achermann, 1999, 2000] amalgam of physiological processes that involves reciprocal interactions between numerous brain regions that is coordinated by multiple neurotransmitter/peptide systems [Lin, 2000;Jones, 2004]. Sleep is exhibited by all animals studied thus far [Campbell and Tobler, 1984;Tobler, 2000] and constitutes a significant percentage of a 24 h period [Carskadon and Dement, 2000;Ohayon et al., 2004]. Alternations between sleep and wakefulness are regulated by a combination of circadian (sleep timing) [Edgar, 1995] and homeostatic factors (sleep need) [Borbely, 1982]. The homeostatic regulation of sleep is based on the observation that following sleep deprivation (SD), there is an increase in sleep time and intensity that is proportional to the sleep lost, suggesting that a need for sleep accumulates during waking in both rodents and humans [Tobler and Borbely, 1986;Riedner et al., 2007;Vyazovskiy et al., 2007]. In humans, SD results in marked cognitive and physiological impairments [Drummond and Brown, 2001;Durmer and Dinges, 2005], while prolonged SD in rats [Rechtschaffen, 1998] and flies [Shaw et al., 2002] is fatal. The biological function(s) of sleep, however, remains poorly characterized, though most agree that sleep serves a restorative function [Benington and Heller, 1995]. Proposed roles for sleep include the maintenance of body temperature [McGinty and Szymusiak, 1990;Wehr, 1992], energy ...

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Proteomic and Immunoblot Analyses of Bartonella quintana Total Membrane Proteins Identify Antigens Recognized by Sera from Infected Patients

Cited by 51 publications

References 48 publications

Proteomic Analysis of Rickettsia parkeri Strain Portsmouth

Proteomic Analysis of Rickettsia parkeri Strain Portsmouth

Detecção de proteínas imunorreativas de Rickettsia sp. cepa Mata Atlântica

Rapid alterations in cortical protein profiles underlie spontaneous sleep and wake bouts

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