Effects of introduced aspartic and glutamic acid residues on the P'1 substrate specificity, pH dependence and stability of carboxypeptidase Y

Carboxypeptidase Y is a serine carboxypeptidase isolated from Saccharomyces cerevisiae with a preference for C-terminal hydrophobic amino acid residues. In order to alter the inherent substrate specificity of CPD-Y into one for basic amino acid residues in P'1, we have introduced Asp and/or Glu residues at a number of selected positions within the S'1 binding site. The effects of these substitutions on the substrate specificity, pH dependence and protein stability have been evaluated. The results presented here demonstrate that it is possible to obtain significant changes in the substrate preference by introducing charged amino acids into the framework provided by an enzyme with a quite different specificity. The introduced acidic amino acid residues provide a marked pH dependence of the (kcat/Km)FA-A-R-OH/(kcat/Km)FA-A-L-OH ratio. The change in stability upon introduction of Asp/Glu residues can be correlated to the difference in the mean buried surface area between the substituted and the substituting amino acid. Thus, the effects of acidic amino acid residues on the protein stability depend upon whether the introduced amino acid protrudes from the solvent accessible surface as defined by the surrounding residues in the wild type enzyme or is submerged below.     

Continuously distributed phase shifts in chirped DFB lasers usingbent waveguides

The authors have implemented chirped distributed feedback (DFB) grating lasers with phase shifts [λ/8, 2λ/8 and 3λ/8] distributed continuously over various axial distances, using their recently proposed method for producing bent optical waveguides. The singlemode stability of the lasers with distributed phase shifts was found to be considerably higher than for lasers with abrupt phase shifts     

Continuously chirped DFB gratings by specially bent waveguides fortunable lasers

We have implemented and studied a new type of tunable multiple-section semiconductor distributed feedback (DFB) laser using tailored chirped DFB gratings. Arbitrarily and continuously chirped DFB gratings are defined by bent waveguides on homogeneous grating fields with ultrahigh spatial precision. The mathematical bending functions are optimized in this case to provide enlarged wavelength tuning ranges. We present the results of model calculations, the technological device realization and experimental results of the DFB laser characterization e.g. a tuning range of 5.5 nm without wavelength gaps and high side mode suppression ratio     

A tractable large-signal dynamic model-application to stronglycoupled distributed feedback lasers

The modulation characteristics of DFB semiconductor lasers have been studied using a transfer matrix method combined with an appropriate rate equation analysis. The model takes into account longitudinal mode spatial hole burning, as well as the nonuniform current injection resulting from the axially varying Fermi voltage, and can be used for the efficient simulation of static, small-signal, and large-signal dynamic properties. The program is applied to the interpretation of experimental data from a strongly coupled InGaAsP/InP DFB laser. The experimental high-frequency properties of this device are well described by the simulations     

Endothelial selectins and vascular cell adhesion molecule-1 promote hematopoietic progenitor homing to bone marrow

The adhesive mechanisms allowing hematopoietic progenitor cells (HPC) homing to the bone marrow (BM) after BM transplantation are poorly understood. We investigated the role of endothelial selectins and vascular cell adhesion molecule-1 (VCAM-1) in this process. Lethally irradiated recipient mice deficient in both P-and E-selectins (P/E-/-), reconstituted with minimal numbers (相似文献   

Expression of beta 2-integrins and L-selectin on polymorphonuclear leukocytes in septic patients

Adhesion molecules on polymorphonuclear leukocytes (PMNL) play an important role in nonspecific defense mechanisms directed at invading microorganisms. When local infection, however, cannot be controlled, a systemic inflammatory response syndrome (SIRS) ensues which may progress to septic shock and multiple organ failure, these being major determinants of the patient's outcome. In the present study, the expression of beta 2-integrins and L-selectin on blood PMNL was measured on subsequent days in patients with sepsis (n = 17) and in healthy volunteers (n = 15). beta 2-Integrins and L-selectin molecules were detected by flow cytometry, using the monoclonal antibodies IB4 (anti-CD18) and Dreg200 (anti-CD62L), respectively. Adhesion molecules were determined at baseline immediately after blood collection and also 45 min after incubation of cells in vitro at body temperature to allow for spontaneous regulation. In addition, PMNL were activated by receptor-dependent and receptor-independent stimuli to characterize stimulus-specific adhesion molecule expression. In parallel with the measurement of adhesion molecules, severity of sepsis was assessed by the Elebute score. The results demonstrate significant differences in the basal, spontaneous and stimulus-induced expression of adhesion molecules between healthy volunteers, survivors (n = 11) and nonsurvivors (n = 6). Moreover, when survivors and nonsurvivors with severe sepsis (Elebute score > 12) were compared, basal expressions of both beta 2-integrins and L-selectin were significantly lower in patients who did not survive. Thus, measurement of adhesion molecules on circulating PMNL may be useful to identify septic patients at high risk for lethal outcome.     

All-atom generalized-ensemble simulations of small proteins

We give an overview of some generalized-ensemble techniques that have proven successful in all-atom simulations of proteins. We show that these techniques enable efficient investigations of secondary structure formation and folding in peptides and small proteins. Results are presented for various alanine-based artificial peptides and a small protein, the 36-residued villin headpiece subdomain (HP-36). Our results indicate that all-atom simulations of proteins may be more restricted by the accuracy of the present energy functions than by the efficiency of the search algorithms.     

核磁共振法确定多孔介质的性能

核磁共振技术(NMR)是一种有力而安全的用于获取介质的空间分辨信息的方法。分析NMR数据的新方法能提供研究各种物理体系的机遇。本文讨论了核磁探针法的实验和分析进展。该方法用于定量评价表征流体和其在多孔介质中的流动行为。这种探针法的核心是辅助函数评价问题的求解,而这类问题的求解决依赖于用B spline表示法对未知函数的规整化和序列评估。