FEMS Microbiol Rev 25:455–501PubMed Wang TW (1980) Amperometric ARN-509 hydrogen electrode. Methods Enzymol 69:409–413. doi:10.​1016/​S0076-6879(80)69040-5 CrossRef Winkler M, Heil B, Heil B, Happe T (2002a) Isolation and molecular characterization

of the [Fe]-hydrogenase from the unicellular green alga Chlorella fusca. Biochim Biophys Acta 1576:330–334PubMed Winkler M, Hemschemeier A, Gotor C, Melis A, Happe T (2002b) [Fe]-hydrogenases in green algae: photo-fermentation and hydrogen evolution under sulfur-deprivation. Int J Hydrogen Energy 27:1431–1439. doi:10.​1016/​S0360-3199(02)00095-2 CrossRef Winkler M, Maeurer C, Hemschemeier A, Happe T (2002c) The isolation of green algal strains with outstanding H2-productivity. In: Miyake J, Igarashi Y, Roegner M (eds) Biohydrogen III. Elsevier Science, Oxford, pp 103–115 LGK 974 Wollman FA (2001) State transitions reveal the dynamics and flexibility of the photosynthetic apparatus. EMBO J 20:3623–3630. doi:10.​1093/​emboj/​20.​14.​3623 CrossRefPubMed Wollman F-A, Delepelaire P (1984) Correlation between changes in light energy distribution and changes in thylakoid membrane polypeptide phosphorylation in Chlamydomonas reinhardtii. J Cell Biol 98:1–7. doi:10.​1083/​jcb.​98.​1.​1 CrossRefPubMed Wykoff DD, Davies JP, Melis A, Grossman AR (1998) The regulation

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“Photo-CIDNP MAS NMR as spectroscopic method Due to small Zeeman splitting and resulting unfavorable Boltzmann distribution, all magnetic resonance methods are intrinsically low in sensitivity. The solid-state photo-CIDNP effect has been shown to be a method to overcome this limitation for magic-angle spinning (MAS) NMR by photochemical production of non-Boltzmann nuclear spin states and to allow for detailed studies of the photochemical machineries of RCs (Zysmilich and McDermott 1994; for reviews: Jeschke and Matysik 2003; Daviso et al. 2008a). Signal enhancement of a factor of about 10,000 for 13C NMR (Fig. 1) has been observed in several RCs (Prakash et al. 2005a, 2006; Roy et al. 2006). The corresponding ratio of the nuclear spin populations of p β/p α = 1.2329 could be expressed in terms of a spin temperature of T S = −0.01146 K. Although temperatures are defined for equilibrium state only, this number may provide an impression about the high degree of spin order obtained. Until now, photo-CIDNP MAS NMR has been measured at fields between 4.

a Linear scale; b semi-log scale Following 400 mg ESL, the BIA 2-005 mean C max values of the test (ESL 400 mg TBM) and reference (ESL 400 mg MF) formulations were 6.4 and 6.3 µg/mL, respectively. The median T max values were 2.0 h for both. Results for the extent of absorption, as determined from mean AUC0–t and AUC0–∞ values, were 105.9 and 106.6 μg h/mL,

respectively, after administration of the Test formulation and 110.3 and 111.1 μg h/mL, respectively, after administration of the reference formulation (Table 1). Table 1 Summary of pharmacokinetics parameters of BIA 2-005 following administration of a single dose of ESL 400 mg and 800 mg TBM and MF formulations Sepantronium solubility dmso BIA 2-005 C max (µg/mL) T max (h) AUC0–t (µg h/mL) AUC0–∞ (µg h/mL) T 1/2 (h) 400 mg ESL (MF)  Geometric mean 6.32 2.0 (0.5–6.0) 110.30 111.13 9.5  Arithmetic mean ± SD 6.46 ± 1.35   112.57 ± 23.01 113.42 ± 23.25 9.6 ± 1.4  CV % 21 59 20 21 15 400 mg ESL (TBM)  Geometric mean 6.39 2.0 (0.5–6.0) 105.85 106.62 9.4  Arithmetic mean ± SD 6.55 ± 1.52   108.22 ± 23.97 109.03 ± 24.25

9.5 ± 1.5  CV % 23 62 22 22 16 800 mg ESL (MF)  Geometric mean 12.95 2.0 (1.0–4.0) 273.47 277.27 11.9  Arithmetic mean ± SD 13.18 ± 2.22   279.04 ± 61.74 282.93 ± 63.32 12.06 ± 1.9  CV % 19 41 22 22 14 800 mg ESL (TBM)  Geometric mean 12.81 1.8 (1.0–6.0) 272.68 277.08 check details 12.2  Arithmetic mean ± SD 12.99 ± 2.56   278.73 ± 60.18 283.39 ± 61.00 12.35 ± 1.7  CV % 17 61 22 22 16 C max, Maximum observed plasma concentration; T max, time to C max (value is median with range); T 1/2, terminal plasma half-life; AUC0–t , area under the concentration-time curve (AUC) from time zero to last Tipifarnib ic50 observable concentration; AUC0–∞, AUC from time zero to infinity; ESL, eslicarbazepine acetate; MF, marketed formulation; TBM, to-be-marketed formulation Following 800 mg ESL, the BIA 2-005 mean C max values of the test (ESL

800 mg TBM) and reference (ESL 800 mg MF) formulations were 12.81 and 12.95 µg/mL, respectively. below The mean t max values were 1.8 and 2.0 h, respectively. Results for the extent of absorption, as determined from mean AUC0–t and AUC0–∞ values, were 272.7 and 277.1 μg h/mL, respectively, after administration of the Test formulation and 273.4 and 277.3 μg h/mL, respectively, after administration of the reference formulation (Table 1). The bioequivalence was evaluated by using the geometric means of C max, AUC0–t and AUC0–∞ values for BIA 2-005. The ratio (test/reference) of each parameter ranged from 96 to 101 % for both dose strengths (Table 2). Following 400 mg ESL, the 90 % confidence intervals for the ratios of C max, AUC0–t and AUC0–∞ were 94–109, 94–98 and 94–98 %, respectively, meeting the predetermined criteria for bioequivalence.

Next, 1 U of RNasin, 2 μl of 100 mM DTT, 1 μl of 10 mM dNTP and 0.5 μl of 200 U/μl MMLV High Performance Reverse Transcriptase (Epicentre, Madison, WI) were added to each RNA/primer mixture and incubated at 37°C for 1 h, followed by heating at 85°C for 10 min to inactivate the selleckchem enzyme and then chilled on ice for at least 1 min. The specific cDNA that we prepared was used in the following quantitative real-time

PCR analysis. The components of real-time PCR were prepared by adding 10 ng of each specific cDNA and 1 μl of a 10 mM primer solution to 2 × Maxima SYBR Green/ROX qPCR Master Mix (Fermentas) and adjusted with ddH2O to a final volume of 20 μl. Cycling conditions were performed using Roche LightCycler 2.0 system (Roche Applied Crenigacestat datasheet Science, Branford, CT) as follows: 95°C for 2 min followed by 40 cycles of 95°C for 30 sec, 50°C for 30 sec and 72°C for 15 sec. Dissociation curves and non-template controls were included to detect any primer dimerization or other artifacts. The mRNA transcript levels were obtained by the method described by Livak and Schmittgen [37]. Fusion protein construction A carboxy terminal 6 × histidine-tagged fusion to STM0551 was constructed by amplifying stm0551 with primers stm0551-TOPO-F and stm0551-TOPO-R using genomic DNA of S. Typhimurium LB5010 as the template. The resulting 316-bp PCR

product was cloned into the pET101/D-TOPO vector (Invitrogen, Carlsbad, CA) giving rise to plasmid pSTM0551-His. This recombinant plasmid was sequenced at the adjacent portion of the cloning site to make sure it was in frame before subsequent transformation step. BL21Star™ (DE3) One Shot® chemically competent E. coli (Invitrogen) cells were transformed with pSTM0551-His. Log phase cultures were

induced to express STM0551-His by adding 1 mM IPTG at 37°C for 4 hr. The STM0551-His fusion protein was further purified by ProBond purification kit (Invitrogen) using the protocol provided by the manufacturer. The protein concentration was determined using the Bradford reagent (Fermentas) [38]. A mutant allele of stm0551 was constructed by site-directed mutagenesis using overlapping-extension PCR of S. Typhimurium LB5010 strain genomic DNA Doxacurium chloride template and mutagenic oligonucleotides E49A-TOPO-F and E49A-TOPO-R [39]. Briefly, STM0551-TOPO-F and E49A-TOPO-R were used to amplify the first DNA fragment using Pfu DNA polymerase (Fermentas). The PCR conditions were: denaturing at 94°C for 3 min followed by 35 cycles of 94°C for 45 sec, 50°C for 45 sec and 72°C for 45 sec. The second DNA fragment was amplified using E49A-TOPO-F and STM0551-TOPO-R with the same ATM Kinase Inhibitor procedure described above. These two DNA fragments were purified by Montage Gel Extraction Kit (Millipore, Billerica, MA).

Multivariate logistic regression analysis showed that current smoking mTOR inhibitor habits were positively associated with albuminuria and inversely associated with a low eGFR. The association between smoking and

GFR was dependent on the number of cigarettes smoked per day. A history of smoking showed a significant inverse association with a low eGFR, but there was no significant association between former smoking status and albuminuria. These data suggest that smoking may increase albuminuria and decrease eGFR, and that albuminuria may be reversed by quitting smoking. Stengel et al. examined data from a non-concurrent cohort study of 9,082 US adults, aged 30-74 years, who participated in the second CYT387 mouse National Health and Nutrition Examination Survey (NHANES II) from 1976 through 1980. The risk of CKD was found to be related to smoking: the Saracatinib ic50 relative risk (RR) in smokers of 1–20 cigarettes a day

versus never-smokers was 1.2 (95 % CI 0.7–2.3), and in smokers of more than 20 cigarettes a day, the RR rose to 2.3 (95 % CI 1.3–4.2). This study suggests that not only quitting smoking, but also cigarette reduction may reduce the development of kidney disease. Shankar et al. performed a longitudinal analysis among 3,392 CKD-free persons at baseline, looking at the incidence of CKD (n = 114) over a 5 year period. Compared to never-smokers, the odds ratio of developing CKD was 1.12 (95 % CI 0.63, 2.00) among former smokers and 1.97 (95 % CI 1.15, 3.36) among current smokers. Haroun et al. performed a community-based, prospective observational study of 20-year duration to examine the association between hypertension

and smoking on the future risk of CKD in 23,534 Tideglusib men and women in a local region. The results showed that current smoking was significantly associated with the risk of developing CKD in both men [hazard ratio 2.4 (1.5–4.0)] and women [hazard ratio 2.9 (1.7–5.0)]. Above all, smoking is a risk factor for the development of CKD and proteinuria, and former smokers may improve albuminuria by quitting smoking compared to current smokers. Therefore, it is recommended to quit smoking. Bibliography 1. Yamagata K, et al. Kidney Int. 2007;71:159–66. (Level 4)   2. Ishizaka N, et al. Hypertens Res. 2008;31:485–92. (Level 4)   3. Stengel B, et al. Epidemiology. 2003;14:479–87. (Level 4)   4. Shankar A, et al. Am J Epidemiol. 2006;164:263–71. (Level 4)   5. Haroun MK, et al. J Am Soc Nephrol. 2003;14:2934–41. (Level 4)   Does increased water intake affect the development of CKD? The effect of increased water intake on the onset and development of CKD is unclear, but dehydration exacerbates kidney function. Clark et al. performed a prospective cohort study in Canada from 2002 to 2008.

J Bacteriol 1999,181(18):5825–5832.PubMed 33. John J, Frech M, Wittinghofer A: Biochemical properties of Ha-ras encoded p21 mutants and mechanism of the autophosphorylation reaction. J Biol Chem 1988,263(24):11792–11799.PubMed 34. Sood P, Lerner CG, Shimamoto T, Lu Q, Inouye M: Characterization of the autophosphorylation of Era, an essential

Escherichia coli GTPase. Mol Microbiol 1994,12(2):201–208.Buparlisib price PubMedCrossRef CB-5083 chemical structure 35. Malen H, Pathak S, Softeland T, de Souza GA, Wiker HG: Definition of novel cell envelope associated proteins in Triton X-114 extracts of Mycobacterium tuberculosis H37Rv . BMC Microbiol 2010, 10:132.PubMedCrossRef 36. Wolfe LM, Mahaffey SB, Kruh NA, Dobos KM: Proteomic definition of the cell wall of Mycobacterium tuberculosis . J Proteome Res 2010,9(11):5816–5826.PubMedCrossRef 37. Datta K, Skidmore JM, Pu K, Maddock JR: The Caulobacter crescentus BAY 1895344 datasheet GTPase CgtAC is required for progression through the cell cycle and for maintaining 50 S ribosomal subunit levels. Mol Microbiol 2004,54(5):1379–1392.PubMedCrossRef 38. Tan J, Jakob U, Bardwell JC: Overexpression of

two different GTPases rescues a null mutation in a heat-induced rRNA methyltransferase. J Bacteriol 2002,184(10):2692–2698.PubMedCrossRef 39. Datta K, Fuentes JL, Maddock JR: The yeast GTPase Mtg2p is required for mitochondrial translation and partially suppresses an rRNA methyltransferase mutant, mrm2. Mol Biol Cell 2005,16(2):954–963.PubMedCrossRef 40. Lapik YR, Misra JM, Lau LF, Pestov DG: Restricting conformational flexibility of the switch II region creates a dominant-inhibitory phenotype in Obg GTPase Nog1. Mol Cell Biol 2007,27(21):7735–7744.PubMedCrossRef 41. Scott JM, Haldenwang WG: Obg, an essential GTP binding protein of Bacillus subtilis , is necessary for stress activation of transcription factor sigma(B). J Bacteriol 1999,181(15):4653–4660.PubMed 42. Parida BK, Douglas

T, Nino C, Dhandayuthapani S: Interactions of anti-sigma factor antagonists click here of Mycobacterium tuberculosis in the yeast two-hybrid system. Tuberculosis (Edinb) 2005,85(5–6):347–355.CrossRef 43. Beaucher J, Rodrigue S, Jacques PE, Smith I, Brzezinski R, Gaudreau L: Novel Mycobacterium tuberculosis anti-sigma factor antagonists control sigmaF activity by distinct mechanisms. Mol Microbiol 2002,45(6):1527–1540.PubMedCrossRef 44. Hecker M, Volker U: General stress response of Bacillus subtilis and other bacteria. Adv Microb Physiol 2001, 44:35–91.PubMedCrossRef 45. Ausubel F, Brent R, Kingston R, Moore D, Seidman J, Smith J, Struhl K: Current Prtocols in Molecular Biology. New York: Wiley; 1989. 46. Stover CK, de la Cruz VF, Fuerst TR, Burlein JE, Benson LA, Bennett LT, Bansal GP, Young JF, Lee MH, Hatfull GF, et al.: New use of BCG for recombinant vaccines. Nature 1991,351(6326):456–460.PubMedCrossRef 47. Mueller-Ortiz SL, Wanger AR, Norris SJ: Mycobacterial protein HbhA binds human complement component C3. Infect Immun 2001,69(12):7501–7511.

38** [6.57] 36.6604 ± 14.39* [8.31] 38.00 ± 11.77* [6.79] Std 84.54 ± 9.39* [5.42] 150.12 ± 16.93** [9.77] 187.20 ± 35.38* [19.96] 171.36 ± 9.10** [5.25] 73.67 ± 9.44* [5.45] * P < 0.05; ** P < 0.01

aIC50 value reported as Conc. ± SD [SEM]; SEM of three independent experiments performed in duplicate bStandard used was trolox cStandard used was ascorbic acid dStandard used was ascorbic acid eStandard used was catechin fStandard used was curcumin Antimitotic activity The levels of the physicochemical parameters of Allium cepa (root number and root length) were recorded after treatment with various drugs at 0, 48 and 72 h and found to cause significant inhibition in the growth of roots in comparison with negative control and positive control. From the observations, buy LY333531 it has been revealed that average root length in (9f) treatment group was decreased significantly (1.06 cm) compared with that of the negative control (3.93 cm) after 72 h of treatment. The root morphology

was nearly normal during the negative control treatment, but at positive control and synthesized compound groups, the roots morphology showed an obvious difference in its appearance in that it turned to slightly yellowish to brownish in colour. Its cytotoxic effect was evident in the form of shortening and decaying of roots, while progressive increases in root length and root numbers were observed in control group. The cytotoxic effect of tested compounds inhibits root growth and mitosis to a significant extent. The compound 9f showed lowest mitotic index (0.41 %) with highest activity buy RXDX-101 Farnesyltransferase among all the treatment groups, and it was also observed that the number of non-dividing cells increased in all treatment groups other than negative control. As there is no antimitotic principle in water, it was considered as negative control. Ethyl methanesulphonate (EMS) was treated as positive control treatment group

and induces DNA damage by a direct mechanism, acting at various sites as a monofunctional ethylating agent of nucleotides (Budavari, 1989; Sega, 1984). Cytogenetic analysis With the objective of investigating the possible mechanism involved in root growth inhibition, cytogenetic analysis was performed (Angayarkanni et al., 2007; Auti et al., 2010; Pavlica et al., 2000). All the tested compounds provoked strong inhibition of the mitotic index, where a statistically significant difference in relation to the control, and the decrease in the mitotic index was positively correlated with the selleckchem electron-releasing group (Table 2). Changes in chromosome and cellular morphology were observed with increasing time. Partial c-mitosis (colchicine-like mitosis) and full c-mitosis, with partially functional spindles and completely normal mitotic phases, were seen in the various cells of the same root tip between 6- and 72-h time period. Cytogenetic alterations were investigated, and the results are depicted in Table 2.

CrossRef 25. Silversmit G, Depla D, Poelman H, Marin GB, De Gryse R: An XPS study on the surface https://www.selleckchem.com/products/BIRB-796-(Doramapimod).html reduction of V 2 O 5 (001) induced by Ar + ion bombardment. Surf Sci 2006, 600:3512–3517.CrossRef 26.

Sun M, Cui X: Anodically grown Si–W codoped TiO 2 nanotubes and its enhanced visible light photoelectrochemical response. Electrochem Commun 2012, 20:133–136.CrossRef 27. Cong Y, Zhang J, Chen F, Anpo M: Synthesis and characterization of nitrogen-doped TiO 2 nanophotocatalyst with high visible light activity. J Phys Chem C 2007, 111:6976–6982.CrossRef 28. Kuo Y-Y, Li T-H, Yao J-N, Lin C-Y, Chien C-H: Hydrothermal crystallization and modification of surface hydroxyl groups of anodized TiO 2 nanotube-arrays for more efficient photoenergy conversion. Electrochim Acta 2012, 78:236–243.CrossRef 29. Kontos AI, Arabatzis IM, Tsoukleris DS, Kontos AG, Bernard MC, Petrakis DE, Falaras P: Efficient photocatalysts by hydrothermal treatment of TiO 2 . Catal Today 2005, 101:275–281.CrossRef

30. Livraghi S, Paganini MC, Giamello Selleckchem MK-8931 E, Selloni A, Di Valentin C, Pacchioni G: Origin of photoactivity of nitrogen-doped titanium dioxide under visible light. J Am Chem Soc 2006, 128:15666–15671.CrossRef 31. Nakano Y, Morikawa T, Ohwaki T, Taga Y: Deep-level optical spectroscopy investigation of N-doped TiO 2 films. Appl Phys Lett 2005, 86:132104–132104. 132103CrossRef 32. Di Valentin C, Pacchioni G, Selloni A, Livraghi S, Giamello E: Characterization of paramagnetic species in N-doped TiO 2 powders by EPR spectroscopy and DFT calculations. J Phys Chem B 2005, 109:11414–11419.CrossRef 33. Nambu A, Graciani J, Rodriguez J, Wu Q, Fujita E, Sanz JF: ZD1839 in vivo N doping of TiO 2 (110): photoemission and density-functional studies. J Chem Physics 2006, 125:094706.CrossRef 34. Kaneco S, Katsumata H, Suzuki T, Ohta K:

Electrochemical reduction of CO 2 to methane at the Cu electrode in methanol with sodium supporting salts and its comparison with other alkaline salts. Energy Fuel 2006, 20:409–414.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions DDL carried out the synthesis, characterization, and photocatalytic reduction experiments. ZHZ participated in the synthesis and SEM characterization experiments. QYL and XDW participated in the XPS and Raman characterizations. MZ and JJY participated in the design and preparation of the manuscript. All authors read and approved the final manuscript.”
“Background One-dimensional zinc oxide (ZnO) nanostructures have attracted considerable attention within the last decade because of unique characteristics such as large aspect ratio, high electron mobility, and electrical and optical anisotropy [1, 2]. Their potential applications in various functional devices, including see more sensors, solar cells, photodetectors, etc., have been noted [3, 4].

Figure 7 Pathology of 125 I implanted pancreatic cancer. Representative HE stained sections from the 0 Gy (A), 2 Gy (B), and 4 Gy (C) groups 28 d after 125I seed implantation were prepared as described in the Materials and Methods section. Tumor volume of pancreatic cancer at 0 and 28 days after 125I seed implantation Representative ultrasonic P005091 datasheet images from 0 and 28 d after implantation of 125I seed in the 0 Gy, 2 Gy, and 4 Gy groups

are shown in Figure 8. Quantitative measurements of tumor volume in the 0 Gy, 2 Gy, and 4 Gy groups are shown in Figure 8C, F, and 8I, respectively. In the 0 Gy group, pancreatic cancer proliferated this website rapidly from 0 d to 28 d after implantation (Figures 8A and 8B). The tumor volume (1240 ± 351 v/mm3) at 28 d was significantly larger than at 0 d (809 ± 261, P < 0.01; Figure 8C). No significant alteration in tumor volume was observed between 0 d and 28 d in the 2 Gy group (Figures 8D and 8E). There was no statistical difference in the tumor volume between 0 d and 28 d in the 2 Gy group (750 ± 300 vs. 830 ± 221, P > 0.05; Figure 8F). More importantly, the 4 Gy group demonstrated that the treatment effectively

eliminated the tumor (Figures 8D and 8E). The tumor volume decreased dramatically, from 845 ± 332 at 0 d to 569 ± 121 at 28 d (P < 0.01; Figure 8I). These results suggest that 125I seed implantation inhibits tumor growth and reduces tumor volume, with 4 Gy being more effective than 2 Gy. Figure 8 Tumor volume 0 and 28 d after 125

I seed implantation. The upper, middle, and lower panels show find more representative ultrasound images from 0 Gy (upper), 2 Gy (middle), and 4 Gy (lower) groups 0 and 28 d post 125I seed implantation. *P < 0.05 compared with 0 d post-implantation; Δ P > 0.05 compared with 0 d post-implantation. Discussion Epigenetic changes in cells are closely linked to tumor occurrence, progression and metastases. DNA methylation is a crucially important epigenetic alteration by which the tumor suppressor gene expression and cell cycle regulation may be substantially altered. Three different DNMTs, specifically DNMT1, DNMT3a and DNMT3b, have critical roles Niclosamide in establishing and maintaining DNA methylation. Many chemotherapeutic agents exert their antitumor effects by inducing apoptosis in cancer cells. The purpose of this study is to investigate whether 125I seed irradiation significantly influences the expression of DNA methyltransferases, promote the cell apoptosis and inhibit the pancreatic cancer growth. SW-1990 pancreatic cancer cells were cultured ex vivo and implanted into the pancreas to create the animal model. The 125I seed irradiation induced apoptosis in SW-1990 cells. Likewise, large numbers of apoptotic cells were present in pancreatic cancer receiving 125I seeds implantation. Irradiation-induced apoptosis became more obvious when the radiation dose increased from 2 Gy to 4 Gy.

Importantly, V110A corresponds Poziotinib molecular weight to the V109A substitution within F. tularensis IglA, which rendered F. tularensis unable to escape from phagosomes, grow within host cells and to cause disease in mice [6]. By combining two or more of the substitutions that had a negative impact on VipB binding, an additive effect was observed. Thus, the double mutants V110A/L113A and D104A/V106A, the triple mutant D104A/V106A/V110A and the quadruple mutant D104A/V106A/V110A/L113A were all essentially unable to bind VipB and produced βR428 clinical trial -galactosidase levels similar to the negative vector control (Figure 2A). Importantly, all VipA mutant alleles were produced at similar

levels in the B2H-reporter strain KDZif1ΔZ, which rules out the possibility that variations in protein levels may account for the differences in VipB-binding (Figure 2B). VipA mutants that appeared not to bind VipB showed marked VipB instability and essentially no protein was detected by Western blot analysis (Figure 2B). Figure 1 Alanine point mutants generated within α-helix 2 of VipA. Shown is the amino acid sequence of residues 103–127 predicted to form α-helix 2 within VipA of V. cholerae strain A1552 as well as the Adriamycin homologous region within IglA of F. tularensis LVS, according to Psipred (http://​bioinf.​cs.​ucl.​ac.​uk/​psipred/​). A

deletion within the first part (Δ104-113) of the α-helix abolishes VipA’s ability to bind to VipB in both B2H and Y2H systems (−), while deletions within the second part (Δ114-123) results in Glycogen branching enzyme a VipA variant that retains VipB binding in the Y2H system, but not in the B2H system (+/−). Amino acids that were replaced with alanine in VipA are indicated by closed triangles. Residues in F. tularensis IglA that

previously were mutated and shown to contribute to efficient IglB binding are indicated also by closed triangles [6]. Figure 2 Bacterial two-hybrid analysis of protein-protein interactions involving VipA and VipB. (A) Contact between VipB and wild-type or mutant VipA, fused to Zif and to the ω subunit of E. coli RNAP respectively, induces transcription from the lacZ promoter of the E. coli reporter strain KDZif1ΔZ, resulting in β-galactosidase activity. As a positive control, MglA-Zif and SspA-ω was used while the negative control corresponds to empty vectors. Shown is the mean β-galactosidase activity ± standard deviation in Miller units produced from 3 independent experiments where two independent transformants were tested on each occasion. Data was subjected to a student’s 2-sided t-test to determine whether the β-galactosidase activity produced by a VipA mutant was significantly different from that of wild-type VipA (*, P < 0.05; ***, P < 0.001).

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