TaNBR1, a Novel Wheat NBR1-like Domain Gene Negatively Regulates Drought Stress Tolerance in Transgenic Arabidopsis

Drought stress is an important factor that severely affects crop yield and quality. Autophagy has a crucial role in the responses to abiotic stresses. In this study, we explore TaNBR1 in response to drought stress. Expression of the TaNBR1 gene was strongly induced by NaCl, PEG, and abscisic acid treatments. The TaNBR1 protein is localized in the Golgi apparatus and autophagosome. Transgenic Arabidopsis plants overexpressing TaNBR1 exhibited reduced drought tolerance. When subjected to drought stress, compared to the wild-type (WT) lines, the transgenic overexpressing TaNBR1 plants had a lower seed germination rate, relative water content, proline content, and reduced accumulation of antioxidant enzymes, i.e., superoxide dismutase, peroxidase, and catalase, as well as higher chlorophyll losses, malondialdehyde contents, and water loss. The transgenic plants overexpressing TaNBR1 produced much shorter roots in response to mannitol stress, in comparison to the WT plants, and they exhibited greater sensitivity to abscisic acid treatment. The expression levels of the genes related to stress in the transgenic plants were affected in response to drought stress. Our results indicate that TaNBR1 negatively regulates drought stress responses by affecting the expression of stress-related genes in Arabidopsis.     

Intramolecular Interaction with the E6 Region Stabilizes the Closed Conformation of the N-SH2 Domain and Concurs with the Self-Inhibitory Docking in Downregulating the Activity of the SHP2 Tyrosine Phosphatase: A Molecular Dynamics Study

The localization and activity of the SHP2 tyrosine phosphatase across different cellular compartments to the target substrates are steered by the binding of phosphotyrosine (pY) peptides to the tandem SH2 domains. The most N-terminal domain (N-SH2) can also keep the enzyme inactive by intramolecular occlusion of the catalytic site. Enzyme activity can be recovered by an allosteric disruption of this self-inhibitory docking upon the binding of pY peptides to the N-SH2 domain. Prior to this, the N-SH2 domain must abandon the closed conformation because it impedes the access of pY peptides to the binding cleft. Although it cooperates with the self-inhibitory docking in the negative regulation of the phosphatase activity, the structural determinants of the stability of the closed conformation in the self-inhibited phosphatase are still elusive. To address this issue, a molecular dynamics simulation study is carried out. It is shown that the closed conformation is stabilized by the interaction of the N-SH2 domain with a conserved peptide portion in the region encoded by PTPN11 exon 6 (E6).     

Antibody Therapies Targeting Complex Membrane Proteins

In analyses of protein families that may serve as drug targets, membrane-associated G-protein-coupled receptors (GPCRs) dominate, followed by ion channels, transporters, and—to a lesser extent—membrane-bound enzymes. However, various challenges put such membrane proteins among key groups of underutilized opportunities for the application of therapeutic antibodies. Antibodies hold the promise of exquisite specificity, as they are able to target even specific conformations of a particular membrane protein, as well as adaptability through engineering into various antibody formats. However, the ease of raising and isolating specific, effective antibodies targeting membrane proteins depends on many factors. In particular, the generation of specific antibodies is easier when targeting larger, simpler, extracellular domains with greater uniqueness of amino acid sequence. The rareness of such ideal conditions is illustrated by the limited number of approved biologics for targeting GPCRs and other complex membrane proteins. Challenges in developing antibodies to complex membrane proteins such as GPCRs, ion channels, transporters, and membrane-bound enzymes can be addressed by the design of the antigen, antibody-generation strategies, lead optimization technologies, and antibody modalities. A better understanding of the membrane proteins being targeted would facilitate mechanism-based drug discovery. This review describes the advantages and challenges of targeting complex membrane proteins with antibodies and discusses the preparation of membrane protein antigens and antibody generation, illustrated by select examples of success.     

Molecular recognition of DNA–protein complexes: A straightforward method combining scanning force and fluorescence microscopy

Combining scanning force and fluorescent microscopy allows simultaneous identification of labeled biomolecules and analysis of their nanometer level architectural arrangement. Fluorescent polystyrene nano-spheres were used as reliable objects for alignment of optical and topographic images. This allowed the precise localization of different fluorescence particles within complex molecular assemblies whose structure was mapped in nanometer detail topography. Our experiments reveal the versatility of this method for analysis of proteins and protein–DNA complexes.     

An in-vitro method for prediction of the digestible crude protein content in pig feeds

An in-vitro method using commercially available enzymes for the prediction of the in-vivo digestible crude protein content was developed and tested on seven feedstuffs and 16 diets for pigs. Fat-extracted feed samples were consecutively incubated with pepsin at pH 1 and pancreatin at pH 6.8. From the nitrogen content of the feed sample and of the residue after incubation the in-vitro digestibility of the crude protein was calculated. Using the linear regression of in-vitro on in-vivo digestible crude protein of samples obtained in feeding trials, the in-vivo digestible crude protein content (DXP_p) in g kg^?1 dry matter could be predicted. For feedstuffs and diets the correlation was 0.99 and 0.95 and the residual standard deviation 17 and 6 g kg^?1 dry matter, respectively. In a similar procedure the nitrogen solubilised during incubation with enzymes was analysed. The regression value was similar to that of the original procedure. However, this procedure was abandoned because it was more laborious.     

Effects of germination on the chemical composition of Glycine and Phaseolus beans

The effects of germination on total protein and amino acids, and phytic acid of beans of Glycine max L, Glycine hispida L, Phaseolus radiatus L and Phaseolus angularis L were studied. The increase in total crude protein content was > 21% for Glycine beans and 8–15% for Phaseolus beans. There was a marked increase in the total essential amino acids of Glycine max (76%) and Phaseolus radiatus (52%). A smaller increase was observed for Phaseolus angularis (25%) and Glycine hispida (3%). The phytic acid contents of the beans were drastically reduced (< 0.2%), mainly due to leaching into the soak water. Total ash content showed a decrease, too, also due to leaching; the loss of potassium was very high whereas losses of the divalent metals, calcium, iron and magnesium, were only moderate, probably because of the ability of divalent cations to bind to protein to form proteincation-phytate complexes and also because divalent salts of phytic acid are insoluble at moderate to high pHs.     

牛血清蛋白在亲水硅片表面吸附的原子力显微成像

本文用原子力显微术（AFM）研究了牛血清白蛋白（BSA）在亲水硅片表面的吸附，硅片表面经亲水处理后，将牛血清蛋白（BSA）吸附在表面，采用轻敲模式，可获得清晰的AFM图像，牛血清蛋白（BSA）的AFM图像表明：BSA在亲水硅片表面是单分子，水平吸附在硅片表面，且吸颗粒状；1mg/ml的BSA在吸附30min后为饱和吸附。BSA到达硅表面后，蛋白中可移动的带正电荷的基团可以趋向亲水表面，使BSA与硅表面的静电相互作用由斥力变为吸引力，BSA可以稳定地吸附在亲水硅片表面。     

一种区域分离方法及其在米粒检测中的应用

针对计算机视觉技术中米粒靠接在一起不利于大米的分级检测,采用图像处理分离靠接在一起的米粒融合区域,提出了基于扫描式区域增长的区域分离方法。该方法充分利用了单个米粒图像区域为凸性区域这一几何特点,通过区域增长的方法在图像中寻找由凸性米粒靠接在一起所形成的融合凹性区,并从凹处将融合区域分离开来。由于利用了米粒区域的几何特征来分离融合米粒区域,因此分离工作的自适应性强。大米垩白粒率的检测实验结果表明：该方法分离融合米粒区域的成功率达到了100％,而且检测结果与人工检测结果基本一致。     

One‐Step Formulation of Protein Microparticles with Tailored Properties: Hard Templating at Soft Conditions

Formulation of therapeutic proteins into particulate forms is a main strategy for site‐specific and prolonged protein delivery as well as for protection against degradation. Precise control over protein particle size, dispersity, purity, as well as mild preparation conditions and minimal processing steps are highly desirable. It is, however, hard to fit all these criteria with conventional preparation techniques. Here a one‐step hard‐templating synthesis of microparticles composed of functional, non‐denatured protein is reported. The method is based on filling porous CaCO₃ microtemplates with the protein near to its isoelectric point (pI) followed by pH‐ or EDTA‐mediated dissolution of the tempplates. In principle, a wide variety of proteins can be converted into microparticles using this approach. The main requirement is an overlap of the protein insolubility and a template solubility for a certain parameter (here pH or EDTA). Here the formulation of insulin particles is studied in detail and it is shown that particles consisting of high molecular weight protein (catalase) can also be prepared. In this context, the synthesis of CaCO₃ templates with controlled size, the mechanism of the protein microparticle formation and mechanical properties of the microparticles are discussed. For the first time, the fabrication of mesoporous monodispersed CaCO₃ microtemplates with identical porocity but tuned diameter from 3 to 20 μm is demonstrated. The protein particle diameter can be adjusted by choosing the appropriate template size that is critical for successful pulmonary delivery of insulin. As a first step towards insulin delivery, the in vitro release of insulin at physiological conditions is studied.     

5种稻田突眼隐翅虫触角感器的超微结构研究

本文利用扫描电镜对小黑突眼隐翅虫Stenus melanarius、瘦突眼隐翅虫S.tenuipes、黑胫足突眼隐翅虫S.macies、虎突眼隐翅虫S.cicindeloides和阑氏突眼隐翅虫S.lewisius等5种稻田突眼隐翅虫雌、雄成虫的触角感器进行了观察和比较研究.结果表明,5种突眼隐翅虫的触角感器类型基本相同,均有5种感器,其中毛形感器和锥形感器又分为3个亚型:即B?hm氏鬃毛、毛形感器(毛形感器1、毛形感器2、毛形感器3)、刺形感器、锥形感器(锥形感器1、锥形感器2、锥形感器3)和栓锥形感器,同种感器形态相似;各种感器在触角上的分布相对稳定,具有一定规律;感器的分布、数量在种间及雌、雄两性间无显著差异.因此,触角感器可能不宜作为突眼隐翅虫种类鉴定的依据之一.