Binding Study of Phenformin with Bovine Serum Album Using a Combined Technique of Equilibrium Dialysis with Flow-Injection Chemiluminescence Detection

ABSTRACT

The interaction between phenformin hydrochloride and bovine serum albumin (BSA) was investigated by the methods of chemiluminescence combined with equilibrium dialysis technique. A novel N-bromosuccinimide (NBS)–eosin Y (EY) chemiluminescence (CL) method was established for the determination of phenformin. The mechanism of this chemiluminescence system was proposed. Optimization studies were performed to determine the phenformin. Under the optimal conditions, the CL intensity was linear for a phenformin concentration over the range of 4.6 × 10^?8 to 5.0 × 10^?5 g/mL. The detection limit was 1.5 × 10^?8 g/mL. The data obtained by the present equilibrium dialysis–CL system were analyzed using the Klotz plot and the Scatchard analysis. The results showed that the Klotz plot and the Scatchard plot are linear with good correlation coefficient, indicating that the phenformin has only one type of binding site on BSA. The binding parameters were the number of the binding sites n (1.02) and the estimated association constant K (2.66 × 10⁴ L/mol). The chemiluminescence system combined with equilibrium dialysis developed in this work demonstrated its use for determination of interaction between drug and protein by using relatively simple instrument.     

Recognition and elimination of diversified pathogens in insect defense systems

Summary The elimination of infectious non-self by the host defense systems of multicellular organisms requires a variety of recognition and effector molecules. The diversity is generated in somatic cells or encoded in the germ-line. In adaptive immunity in jawed vertebrates, the diversity of immunoglobulins and antigen receptors is generated by gene rearrangements in somatic cells. In innate immunity, various effector molecules and pattern recognition receptors, such as antimicrobial peptides and peptidoglycan recognition proteins, are encoded in the germ-line of multicellular organisms, including insects and jawed vertebrates. In the present review, we discuss how insect host defense systems recognize and eliminate a multitude of microbes via germ-line-encoded molecules, including recent findings that a Drosophila member of the immunoglobulin superfamily is extensively diversified by alternative splicing in somatic immune cells and participates in the elimination of bacteria.     

Statistical Thermodynamic Properties of Linear Protein Solutions

利用统计力学理论结合格点模型,讨论了链状蛋白质分子溶液的热力学性质. 结果表明,对于稀溶液来说,溶液的吉布斯函数随蛋白质浓度的增加而降低,蛋白质分子化学势随其浓度增加而升高. 还分析了蛋白质分子链长及温度对溶液吉布斯函数和蛋白质分子化学势的影响. 并且计算讨论了几种第一类抗冻蛋白的化学势.     

Modifications to functional and biological properties of proteins of cowpea pulse crop by ultrasound-assisted extraction

Natural resource depletion, negative environmental effects and the challenge to secure global food security led to the establishment of the Sustainable Development Goals (SDGs). In need to explore underutilized sustainable protein sources, this study aims at isolating protein from cowpea by ultrasound-assisted extraction (UAE), where the techno-functional characteristics of the protein isolates were studied at different sonication conditions i.e., 100 W and 200 W at processing times ranging from 5 to 20 min. The US at 200 W-10 min produced the optimal results for all properties. In this process combination, there was an increase in protein yield, solubility, water-holding capacity, foaming capacity and stability, emulsion activity and stability, zeta-potential, and in-vitro protein digestibility from 31.78% to 58.96%, 57.26% to 68.85%, 3.06 g/g to 3.68 g/g 70.64% to 83.74%, 30.76% to 60.01%, 47.48% to 64.26%, 56.59% to 87.71%, –32.9 mV to −44.2 mV and 88.27% to 89.99%, respectively and particle size dropped from 763 nm to 559 nm in comparison to control. The microstructure and secondary-structure alterations of proteins caused by sonication were validated by SEM images, SDS-PAGE, and FTIR analyses. Sonication leads to acoustic cavitation and penetrate the cell walls, improving extraction from the solid to liquid phase. After sonication, the hydrophobic protein groups were exposed and proteins were partially denatured which increased its functionality. The findings demonstrated that UAE of cowpea protein improved yield, modify characteristics to fit the needs of the food industry, and contribute to achieving SDGs 2, 3, 7, 12, and 13.     

磷脂酶A2家族的功能性分类研究

采用“结合强度指纹图谱分析”方法, 通过对多重分子对接得到的作用强度数据进行聚类矩阵分析对蛋白质进行功能分类. 着重研究了磷脂酶A₂家族基于抑制剂作用强度的功能分类, 并且与基于序列的聚类结果进行比较, 成功地解决了序列比对方法不能处理的远源蛋白(cPLA₂)的分类问题.     

用单分子技术研究Sso7d与DNA的相互作用

为了维持基因的稳定性,每种生物体都含有一套独特的染色质蛋白来保护脱氧核糖核酸(DNA)的结构,观察染色质蛋白对DNA结构的作用过程和结果,可以帮助人们了解这些蛋白的具体功能和作用机理.硫化叶菌是一种能在高温下存活的古细菌,Sso7d是硫化叶菌的一种染色质蛋白.深入地了解Sso7d和DNA链的相互作用,有助于解释硫化叶菌的DNA为何能在高温环境下保持活性,本文通过原子力显微镜(AFM)和磁镊两种单分子操作手段,研究了Sso7d与DNA的相互作用.AFM的实验结果给出了Sso7d与DNA的作用过程:结合Sso7d后,DNA首先发生弯折,然后出现loop结构,最终DNA会团聚为致密的核结构.利用磁镊装置测量了Sso7d的结合对打开DNA双链的影响,实验结果表明Sso7d的结合导致打开DNA双链的力的增大,经过数据分析,计算出Sso7d与DNA结合的结合能?G=3.1k_BT,平均每5.5个碱基对(bp)结合一个Sso7d,较高的结合密度和较大的结合能,两方面的作用结果,解释了Sso7d能够稳定DNA结构的原因.     

聚乳酸-聚半胱氨酸混纺纤维毡用于谷胱甘肽S-转移酶的捕获和洗脱

本文研究一种分离纯化GST（谷胱甘肽S-转移酶）的方法：将嵌段聚合物聚左旋乳酸-聚半胱氨酸（PLLA—PCys）电纺丝制成超细纤维,将还原型谷胱甘肽（GSH）通过二硫键偶联到纤维上,再将纤维填充玻璃管,制成吸附柱;含有GST的混合溶液通过吸附柱时,GST被纤维捕获;再用含GSH的溶液流过吸附柱,GST被洗脱下来,经冷冻干燥获得GST纯品．用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳进行检测．结果表明,键合了GSH的PLLA—PCys混纺纤维毡能够从蛋白混合溶液中专一性地捕获GST,被捕获的GST可以有效地回收．所以,此种键合了GSH的电纺丝纤维可望用于GST或以GST为标签的蛋白的分离、纯化以及检测．     

Investigating Membrane-Mediated Antimicrobial Peptide Interactions with Synchrotron Radiation Far-Infrared Spectroscopy

Synchrotron radiation-based Fourier transform infrared spectroscopy enables access to vibrational information from mid over far infrared to even terahertz domains. This information may prove critical for the elucidation of fundamental bio-molecular phenomena including folding-mediated innate host defence mechanisms. Antimicrobial peptides (AMPs) represent one of such phenomena. These are major effector molecules of the innate immune system, which favour attack on microbial membranes. AMPs recognise and bind to the membranes whereupon they assemble into pores or channels destabilising the membranes leading to cell death. However, specific molecular interactions responsible for antimicrobial activities have yet to be fully understood. Herein we probe such interactions by assessing molecular specific variations in the near-THz 400–40 cm⁻¹ range for defined helical AMP templates in reconstituted phospholipid membranes. In particular, we show that a temperature-dependent spectroscopic analysis, supported by 2D correlative tools, provides direct evidence for the membrane-induced and folding-mediated activity of AMPs. The far-FTIR study offers a direct and information-rich probe of membrane-related antimicrobial interactions.     

Effects of aging and maternal protein restriction on the muscle fibers morphology and neuromuscular junctions of rats after nutritional recovery

Changes in the nutritional status of mothers may predispose their offspring to neuromuscular disorders in the long term. This study evaluated the effects of maternal protein restriction during pregnancy and lactation on the muscle fibers and neuromuscular junctions (NMJs) of the soleus muscle in the offspring of rats at 365 days of age that had undergone nutritional recovery. Wistar rats were divided into two groups: control (CG) – the offspring of mothers fed a normal protein diet (17%) and restricted (RG) – offspring of mothers fed a low protein diet (6%). After lactation, the male pups received standard chow ad libitum. At 365 days, samples of soleus muscle were collected for muscle fiber analysis (HE staining, NADH-TR reaction and ultrastructure), intramuscular collagen quantification (picrosirius red staining) and NMJs analysis (non-specific esterase technique). The cross-sectional area of type I fibers was reduced by 20% and type IIa fibers by 5% while type IIb fibers increased by 5% in the RG compared to the CG. The percentage of intramuscular collagen was 19% lower in the RG. Disorganization of the myofibrils and Z line was observed, with the presence of clusters of mitochondria in both groups. Regarding the NMJs, in the RG there was a reduction of 10% in the area and 17% in the small diameter and an increase of 7% in the large diameter. The results indicate that the effects of maternal protein restriction on muscle fibers and NMJs seem to be long-lasting and irreversible.     

Effects of high hydrostatic pressure on secondary structure and emulsifying behavior of sweet potato protein

In this study, secondary structures of sweet potato protein (SPP) after high hydrostatic pressure (HHP) treatment (200–600?MPa) were evaluated and emulsifying properties of emulsions with HHP-treated SPP solutions in different pH values (3, 6, and 9) were investigated. Circular dichroism analysis confirmed the modification of the SPP secondary structure. Surface hydrophobicity increased at pH 3 and decreased at 6 and 9. Emulsifying activity index at pH 6 increased with an increase in pressure, whereas emulsifying stability index increased at pH 6 and 9. Oil droplet sizes decreased, while volume frequency distribution of the smaller droplets increased at pH 3 and 6 with the HHP treatment. Emulsion viscosity increased at pH 6 and 9 and pseudo-plastic flow behaviors were not altered for all emulsions produced with HHP-treated SPP. These results suggested that HHP could modify the SPP structure for better emulsifying properties, which could increase the use of SPP emulsion in the food industry.