The availability of the small molecular lead-compound library and the modeled 3D target structure makes it possible mTOR inhibitor to use SBVS to screen out a limited number of promising candidates

that can interrupt the TCS signal transduction by interacting with the HKs substrate of S. pneumoniae. HKs, as novel antibacterial targets, have attracted many attentions due to their essentiality in the viability of microbes and their deficiency in animals. HKs are involved in the regulation of bacterial growth and virulence in many bacterial species. Previously, a HK named VicK has been used to screen lead compound inhibitors in B. subtilis and S. epidermidis. We here for the first time obtained 105 candidate chemical compounds directly aiming at S. pneumoniae VicK by screening 200,000 possible compounds in silico. Compounds that can bind to the purified target protein VicK’ and compete with its substrate ATP were further verified by in vitro and in vivo antibacterial assays. Eventually, we obtained 6 compounds with antibacterial activity that may be used as novel drug leads. Commonly, the response

regulator YycF and the histidine kinase YycG are the only essential TCS for viability in B. subtilis and Go6983 datasheet S. aureus [10, 12]. In S. pneumoniae, the VicR/K TCS regulates the expression of several critical genes, such as those encoding surface proteins and virulence factors [21, 33]. However, only the response regulator VicR was found to be essential [20, 34]. The signal transduction of VicK was possibly bypassed by other TCS HKs [35]. VicK has conserved ATP-dependent HATPase_c domains accounting for autophosphorylation. Even non-cognate HKs from other bacteria can ABT-737 solubility dmso phosphorylate the purified VicR from S. pneumoniae [18]. In a previous study [36],

the MIC values of the lead compounds PAK6 screened out by SBVS targeting the YycG of S. epidermidis were almost equal to the corresponding IC50 (for YycG’) values, with a correlation coefficient of 0.959, which suggested that inhibition of 50% the YycG protein activity would interfere with the growth of S. epidermidis. If this case is true in S. pneumoniae, the result that the MIC values of the lead-compounds were far less than the corresponding IC50 values may be explained as bypass effects of these compounds on other HKs. In a word, these lead compounds are most likely having a “”cross-inhibition”" on other HKs in S. pneumoniae, which can enhance their antibacterial effects, although they were not verified in this study. Although the VicK protein in S. pneumoniae can be homologous to YycG in other Gram-positive strains, such as S. epidermidis, Enterococcus faecalis and S. aureus, different strains generally have different characteristics of the HATPase_c domain structure of HKs. These characteristics will determine the binding specifiCity of the lead compounds screened out by SBVS.