Design and Characterization of a New pVII Combinatorial Phage Display Peptide Library for Protease Substrate Mining Using Factor VII Activating Protease (FSAP) as Model

We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated.     

Comparative study of surface properties of Mg-substituted hydroxyapatite bioceramic microspheres

Mg-substituted hydroxyapatite (HAp) bioceramic microspheres were prepared by spray drying and subsequent processing at 1173, 1273 and 1373 K. Influence of various Mg substitution levels (up to 0.84 ± 0.10 wt%) on physicochemical properties of the HAp bioceramic microspheres was evaluated. Obtained results were used for the elucidation of the compositional and structural characteristics of the microspheres in conjunction with adsorption of protein, namely, bovine serum albumin (BSA). The primary difference among the microspheres processed at various temperature was the presence or absence of the micropores (<2 nm in diameter) and mesopores (between 2 and 50 nm). Presence of the micro- and mesopores resulted in higher specific surface area (SSA), enhanced solubility, i.e., ion release, and, accordingly, increase in the amount of BSA adsorbed on the microspheres. Furthermore, the BSA adsorption capacity of the microspheres decreased with increasing Mg content despite of higher SSA.     

Parallel color-coding

We present new parallelization and memory-reducing strategies for the graph-theoretic color-coding approximation technique, with applications to biological network analysis. Color-coding is a technique that gives fixed parameter tractable algorithms for several well-known NP-hard optimization problems. In this work, by efficiently parallelizing steps in color-coding, we create two new biological protein interaction network analysis tools: Fascia for subgraph counting and motif finding and FastPath for signaling pathway detection. We demonstrate considerable speedup over prior work, and the optimizations introduced in this paper can also be used for other problems where color-coding is applicable.     

Protein-based nanomaterials and nanosystems for biomedical applications: A review

Advanced protein-based nanomaterials and nanosystems (PNNS) have attracted considerable scientific interest in recent decades due to their potential in bio-applications. Nowadays, the constructed PNNS exhibit different properties for various special applications based on the characteristics of different proteins. Herein, in this review article, a systematic summary and discussion focusing on designing multi-functional PNNS are presented. The latest developments in unique synthesis strategies and detailed classification of PNNS are reviewed. The functions of proteins in PNNS for biomedical applications, such as targeting proteins, carriers, enzymes, and fluorescent indicators, are summarized. Finally, the challenges and forward-looking perspectives of PNNS research are provided.     

Protein–Energy Malnutrition: A Risk Factor for Various Ailments

The wheel of industrialization that spun throughout the last century resulted in urbanization coupled with modifications in lifestyles and dietary habits. However, the communities living in developing economies are facing many problems related to their diet and health. Amongst, the prevalence of nutritional problems especially protein–energy malnutrition (PEM) and micronutrients deficiencies are the rising issues. Moreover, the immunity or susceptibility to infect-parasitic diseases is also directly linked with the nutritional status of the host. Likewise, disease-related malnutrition that includes an inflammatory component is commonly observed in clinical practice thus affecting the quality of life. The PEM is treatable but early detection is a key for its appropriate management. However, controlling the menace of PEM requires an aggressive partnership between the physician and the dietitian. This review mainly attempts to describe the pathophysiology, prevalence and consequences of PEM and aims to highlight the importance of this clinical syndrome and the recent growth in our understanding of the processes behind its development. Some management strategies/remedies to overcome PEM are also the limelight of the article. In the nutshell, early recognition, prompt management, and robust follow up are critical for best outcomes in preventing and treating PEM.     

Silica-supported carboxylated cellulose nanofibers for effective lysozyme adsorption: Effect of macropore size

Engineering hierarchical macro/mesoporous structures that offer an abundance of accessible binding sites are highly desirable in protein adsorption processes. However, numerous significant challenges remain. Herein, cellulose nanofiber (CNF)-loaded macroporous silica (CNF-MPS) particles were successfully synthesized with a high degree of accessible binding sites by tuning the macropore size of the silica particles and loading a highly carboxylated CNF via smart and rational design. The as-prepared CNF-MPS particles exhibited a high negative charge (~−59 mV) and excellent protein adsorption ability (>1000 mg/g) in <5 min. Furthermore, tuning the macropore size influenced the CNF deposition either to the external surface or penetrating within the pores. As a result, the optimum macropore successfully enhances the adsorption capability to >1500 mg/g as a result of improved interconnectivity between the channels. Here exposed macropores of >100 nm allows ingress of protein to the interior structure that houses an abundance of binding sites comprising the dispersed CNF. Additionally, the adsorption kinetics, thermodynamics, and isothermal parameters were studied to analyze the mechanism of lysozyme adsorption. The adsorption process is confirmed to occur spontaneously at any temperature with a pseudo-second-order model describing the kinetic model, and CNF deposition affecting the heterogeneity of the binding sites.     

全自动凯氏定氮仪测定蛋白质(以N计)不确定度分析与探讨

目的:用全自动凯氏定氮仪测定蛋白质(以N计)不确定度进行分析与探讨。方法 :采用GB 5009.5-2016第一法凯氏定氮法测定蛋白质(以N计)含量,建立数学模型,通过对各个不确定度分量的计算合成,找出影响测量不确定度的因素,最终得出样品中蛋白质(以N计)含量的扩展不确定度与合成标准不确定度。结果:蛋白质(以N计)含量测定的扩展不确定度为(20.93±0.25)%,k=2。结论:采用JJF 1059.1-2012《测量不确定度评定与表示》,对不确定度进行评定,确保蛋白质(以N计)含量结果的准确性。     

Rational Design of Programmable Monodisperse Semi-Synthetic Protein Nanomaterials Containing Engineered Disulfide Functionality**

The reversible nature of disulfide functionality has been exploited to design intelligent materials such as nanocapsules, micelles, vesicles, inorganic nanoparticles, peptide and nucleic acid nanodevices. Herein, we report a new chemical methodology for the construction redox-sensitive protein assemblies using monodisperse facially amphiphilic protein-dendron bioconjugates. The disulfide functionality is strategically placed between the dendron and protein domains. The custom designed bioconjugates self-assembled into nanoscopic objects of a defined size dictated by the nature of dendron domain. The stimuli-responsive behavior of the protein assemblies is demonstrated using a suitable redox trigger.     

Preparative Protein Crystallization

Preparative protein crystallization can possibly replace one or more chromatographic steps in downstream processing. The development of such crystallization processes is demanding: first, promising principal crystallization conditions must be identified; second, details about the process must be defined; and third, the crystals have to be separated from the mother liquor without putting harm of their crystalline integrity. State‐of‐the‐art about these three steps is developing fast by (i) employing new screening methods which are based on fundamental understanding of the interaction of the protein molecules, (ii) application of existing concepts of technical bulk crystallization of small molecules to preparative protein crystallization, and (iii) making available specific gentle separation machinery.