Cysteine-scanning mutagenesis of helix IV and the adjoining loops in the lactose permease of Escherichia coli: Glu126 and Arg144 are essential. off

Cys-scanning mutagenesis has been applied to the remaining 45 residues in lactose permease that have not been mutagenized previously (from Gln100 to Arg144 which comprise helix IV and adjoining loops). Of the 45 single-Cys mutants, 26 accumulate lactose to > 75% of the steady state observed with Cys-less permease, and 14 mutants exhibit lower but significant levels of accumulation (35-65% of Cys-less permease). Permease with Phe140-->Cys or Lys131-->Cys exhibits low activity (15-20% of Cys-less permease), while mutants Gly115-->Cys, Glu126-->Cys and Arg144-->Cys are completely unable to accumulate the dissacharide. However, Cys-less permease with Ala or Pro in place of Gly115 is highly active, and replacement of Lys131 or Phe140 with Cys in wild-type permease has a less deleterious effect on activity. In contrast, mutant Glu126-->Cys or Arg144-->Cys is inactive with respect to both uphill and downhill transport in either Cys-less or wild-type permease. Furthermore, mutants Glu126-->Ala or Gln and Arg144-->Ala or Gln are also inactive in both backgrounds, and activity is not rescued by double neutral replacements or inversion of the charged residues at these positions. Finally, a mutant with Lys in place of Arg144 accumulates lactose to about 25% of the steady state of wild-type, but at a slow rate. Replacement of Glu126 with Asp, in contrast, has relatively little effect on activity. None of the effects can be attributed to decreased expression of the mutants, as judged by immunoblot analysis. Although the activity of most of the single-Cys mutants is unaffected by N-ethylmaleimide, Cys replacement at three positions (Ala127, Val132, or Phe138) renders the permease highly sensitive to alkylation. The results indicate that the cytoplasmic loop between helices IV and V, where insertional mutagenesis has little effect on activity [McKenna, E., et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 11954-11958], contains residues that play an important role in permease activity and that a carboxyl group at position 126 and a positive charge at position 144 are absolutely required.     

On stability analysis of systems featuring a multiplicative combination of nonlinear and linear time‐invariant feedback

We show that sufficient conditions for the finite‐gain stability of certain systems featuring a multiplicative combination of memoryless nonlinear feedback and linear time‐invariant feedback, as encountered in some systems' biology prototype applications, can be obtained by building on the Rantzer multipliers. These results are obtained by observing that for all finite energy signals x, the time‐truncated inner product 〈x, H(x)N(x)〉_T is positive for all T>0 if N belongs to a subclass of positive memoryless monotone nonlinearities and if, in addition, H is a Rantzer multiplier. Copyright © 2011 John Wiley & Sons, Ltd.     

Sulphonic acids doped poly(N-ethyl aniline): A material for humidity sensing application

Substituted polyaniline, poly(N-ethyl aniline) (PNEA), doped with camphor sulphonic acid (CSA) and p-toluene sulphonic acid (p-TSA), was synthesized by single-step chemical polymerization method using ammonium persulphate as an oxidizing agent. This is a single-step polymerization process to synthesize directly the conducting emeraldine salt phase of the polymer using CSA and p-toluene sulphonic acid as dopants. The synthesized polymers were characterized by UV–vis and FTIR spectroscopy, scanning electron microscopy, TGA/SDTA, DSC, and conductivity measurements. This single-step chemical synthesis method offers a polymer having very good physicochemical properties with good electrical conductivity. High temperature conductivity measurements show “thermal activated behavior.” The change in resistance with respect to % relative humidity (% RH) is observed, when pressed pellets of the polymer were exposed to the broad range of humidity (ranging between 20 and 100% RH). POLYM. ENG. SCI., 47:1621–1629, 2007. © 2007 Society of Plastics Engineers     

Cytoplasmic creatine kinases from giant pandas

The muscle and brain creatine kinases of giant panda have been isolated and purified. The purified muscle and brain enzymes (MM and BB) are homogeneous on both the polyacrylamide gel electrophoresis in the presence and absence of SDS. Both enzymes are dimers, consisting of two identical subunits each with a molecular weight of 42,000 daltons. The characteristics of muscle and brain enzymes have been studied, respectively. The hybridized enzyme, MB, was prepared by hybridization of MM and BB. The kinetic parameters of MM, BB and MB were determined, respectively. The results from modification of SH groups show that the SH groups of panda creatine kinases are essential for their activity and among the all SH groups in the enzyme only one per subunit is essential for enzymatic activity.     

Immunoglobulin A antibodies to Helicobacter pylori

Serological testing for immunoglobulin G (IgG) antibodies to Helicobacter pylori has proven useful in supporting the diagnosis of infection with this organism, but the clinical value of IgA antibodies in H. pylori-related gastritis remains controversial. The purpose of our study was to determine the frequency of IgA-positive IgG-negative patients with symptoms of gastrointestinal (GI) disorders, thus assessing the clinical utility of IgA testing for H. pylori-related gastritis. It was found previously that the frequency of infected individuals in this category (IgA positive and IgG negative) is about 2%, but a large number of IgG-negative patients with GI disorders suggestive of H. pylori infection have not been investigated until now.     

NH4Zr2V3O12 proton conductor

NH₄Zr₂V₃O₁₂, a new proton conductor, has been synthesized by flux, melt and hydrothermal methods. The crystals were subjected to X-ray diffraction, differential thermal analysis, infrared spectroscopy and impedance measurements.