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Protein engineering

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https://www.readbyqxmd.com/read/28110414/expression-of-the-citrus-cstip2-1-gene-improves-tobacco-plant-growth-antioxidant-capacity-and-physiological-adaptation-under-stress-conditions
#1
Cristina P S Martins, Diana M Neves, Luciana C Cidade, Amanda F S Mendes, Delmira C Silva, Alex-Alan F Almeida, Mauricio A Coelho-Filho, Abelmon S Gesteira, Walter S Soares-Filho, Marcio G C Costa
Overexpression of the citrus CsTIP2;1 improves plant growth and tolerance to salt and drought stresses by enhancing cell expansion, H 2 O 2 detoxification and stomatal conductance. Tonoplast intrinsic proteins (TIPs) are a subfamily of aquaporins, belonging to the major intrinsic protein family. In a previous study, we have shown that a citrus TIP isoform, CsTIP2;1, is highly expressed in leaves and also transcriptionally regulated in leaves and roots by salt and drought stresses and infection by 'Candidatus Liberibacter asiaticus', the causal agent of the Huanglongbing disease, suggesting its involvement in the regulation of the flow of water and nutrients required during both normal growth and stress conditions...
January 21, 2017: Planta
https://www.readbyqxmd.com/read/28110114/tropoelastin-coated-plla-plga-scaffolds-promote-vascular-network-formation
#2
Shira Landau, Ariel A Szklanny, Giselle C Yeo, Yulia Shandalov, Elena Kosobrodova, Anthony S Weiss, Shulamit Levenberg
The robust repair of large wounds and tissue defects relies on blood flow. This vascularization is the major challenge faced by tissue engineering on the path to forming thick, implantable tissue constructs. Without this vasculature, oxygen and nutrients cannot reach the cells located far from host blood vessels. To make viable constructs, tissue engineering takes advantage of the mechanical properties of synthetic materials, while combining them with ECM proteins to create a natural environment for the tissue-specific cells...
January 11, 2017: Biomaterials
https://www.readbyqxmd.com/read/28110073/sphingosine-1-phosphate-improves-endothelialization-with-reduction-of-thrombosis-in-recellularized-human-umbilical-vein-graft-by-inhibiting-syndecan-1-shedding-in-vitro
#3
Kai Hsia, Ming-Jie Yang, Wei-Min Chen, Chao-Ling Yao, Chih-Hsun Lin, Che-Chuan Loong, Yi-Long Huang, Ya-Ting Lin, Arthur D Lander, Hsinyu Lee, Jen-Her Lu
: Sphingosine-1-phosphate (S1P) has been known to promote endothelial cell (EC) proliferation and protect Syndecan-1 (SDC1) from shedding, thereby maintaining this antithrombotic signal. In the present study, we investigated the effect of S1P in the construction of a functional tissue-engineered blood vessel by using human endothelial cells and decellularized human umbilical vein (DHUV) scaffolds. Both human umbilical vein endothelial cells (HUVEC) and human cord blood derived endothelial progenitor cells (EPC) were seeded onto the scaffold with or without the S1P treatment...
January 18, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28110067/poly-glycerol-sebacate-elastomer-supports-bone-regeneration-by-its-mechanical-properties-being-closer-to-osteoid-tissue-rather-than-to-mature-bone
#4
S H Zaky, K W Lee, J Gao, A Jensen, K Verdelis, Y Wang, A J Almarza, C Sfeir
: Mechanical load influences bone structure and mass. Arguing the importance of load-transduction, we investigated the mechanisms inducing bone formation using an elastomeric substrate. We characterized Poly (glycerol sebacate) (PGS) in-vitro for its mechanical properties, compatibility with osteoprogenitor cells regarding adhesion, proliferation, differentiation under compression versus static cultures and in-vivo for the regeneration of a rabbit ulna critical size defect. The load-transducing properties of PGS were compared in-vitro to a stiffer poly lactic-co- glycolic-acid (PLA/PGA) scaffold of similar porosity and interconnectivity...
January 18, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28109905/cloning-genetic-engineering-and-characterization-of-tmof-expressed-in-saccharomyces-cerevisiae-to-control-larval-mosquitoes
#5
Borovsky Dov, Sabine Nauewelaers, Charles A Powell, Robert G Shatters
Trypsin modulating oostatic factor, a decapaptide isolated from the ovaries of A. aegypti, is the physiological factor that terminates the trypsin biosynthesis after the blood meal. Earlier results obtained from feeding mosquito larvae and injecting female mosquitoes with TMOF show that trypsin biosynthesis and egg development are inhibited, indicating that TMOF traverses the gut epithelial cells and modulates trypsin biosynthesis, making it a potential larvacidal peptide hormone. Therefore, TMOF and TMOF green fluorescent protein (GFP) fusion protein with a trypsin cleavage site, allowing TMOF release in the larval gut, were expressed in S...
January 18, 2017: Journal of Insect Physiology
https://www.readbyqxmd.com/read/28109812/chitosan-as-a-vehicle-for-growth-factor-delivery-various-preparations-and-their-applications-in-bone-tissue-regeneration
#6
REVIEW
Jayachandran Venkatesan, Sukumaran Anil, Se-Kwon Kim, Min Suk Shim
The replacement of conventional autografts and allografts by bone fragments constructed from alternate materials, cells, and molecules (growth factors, drugs, etc.) is an exciting prospect in the field of bone tissue engineering. Bone morphogenetic protein-2 (BMP-2) is a growth factor that has been extensively studied from this point of view. This review analyzes the relevance of chitosan and its derivatives and composites with various materials such as ceramics, heparin, silica, stem cells, titanium implants, etc...
January 18, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28109425/beyond-the-natural-proteome-nondegenerate-saturation-mutagenesis-methodologies-and-advantages
#7
M M Ferreira Amaral, L Frigotto, A V Hine
Beyond the natural proteome, high-throughput mutagenesis offers the protein engineer an opportunity to "tweak" the wild-type activity of a protein to create a recombinant protein with required attributes. Of the various approaches available, saturation mutagenesis is one of the core techniques employed by protein engineers, and in recent times, nondegenerate saturation mutagenesis is emerging as the approach of choice. This review compares the current methodologies available for conducting nondegenerate saturation mutagenesis with traditional, degenerate saturation and briefly outlines the options available for screening the resulting libraries, to discover a novel protein with the required activity and/or specificity...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28107990/combined-use-of-peptide-ion-and-normalized-delta-scores-to-evaluate-milk-authenticity-by-ion-trap-based-proteomics-coupled-with-error-tolerant-searching
#8
Donatella Nardiello, Anna Natale, Carmen Palermo, Maurizio Quinto, Diego Centonze
A fundamental issue in proteomics is the peptide identification by database searching and the assessment of the goodness of fit between experimental and theoretical data. Despite the different number of ways to measure the quality of search results, the definition of a scoring criterion is still highly desirable in ion-trap based proteomics. Indeed, in order to fully take advantage of a low resolution MS/MS dataset, it is essential to strike a balance between greater information capture and reduced number of incorrect peptide assignments...
March 1, 2017: Talanta
https://www.readbyqxmd.com/read/28107808/in-situ-release-of-vegf-enhances-osteogenesis-in-3d-porous-scaffolds-engineered-with-osterix-modified-adipose-derived-stem-cells
#9
Wan-Lin Xu, Hui-Shan Ong, Yun Zhu, Sheng-Wen Liu, Li-Min Liu, Kai-Hua Zhou, Zeng-Qi Xu, Jun Gao, Yan Zhang, Jin-Hai Ye, Wen-Jun Yang
Adipose-derived stem cells (ADSCs) can differentiate into various cell types and thus have great potential for regenerative medicine. Herein, rat ADSCs were isolated; transduced with lentiviruses expressing Osterix (Osx), a transcriptional factor essential for osteogenesis. Osx overexpression up-regulated key osteogenesis-related genes, such as special AT-rich binding protein 2, alkaline phosphatase, osteocalcin and osteopontin, at both the mRNA and protein levels. In addition, mineral nodule formation and alkaline phosphatase activity were enhanced in Osx-overexpressing ADSCs...
January 20, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28107625/gold-nanoshell-mediated-remote-myotube-activation
#10
Attilio Marino, Satoshi Arai, Yanyan Hou, Andrea Degl'Innocenti, Valentina Cappello, Barbara Mazzolai, Young-Tae Chang, Virgilio Mattoli, Madoka Suzuki, Gianni Ciofani
Mild heat stimulation of muscle cells within the physiological range represents an intriguing approach for the modulation of their functions. In this work, photothermal conversion was exploited to remotely stimulate striated muscle cells by using gold nanoshells (NS) in combination with near-infrared (NIR) radiation. Temperature increments of approximately 5°C were recorded by using an intracellular fluorescent molecular thermometer and were demonstrated to efficiently induce myotube contraction. The mechanism at the base of this phenomenon was thoroughly investigated and was observed to be a Ca2+-independent event directly involving actin-myosin interactions...
January 20, 2017: ACS Nano
https://www.readbyqxmd.com/read/28107586/revealing-additional-stereocomplementary-pairs-of-old-yellow-enzymes-by-rational-transfer-of-engineered-residues
#11
Nathalie Nett, Sabine Duewel, Alexandra Annelis Richter, Sabrina Hoebenreich
Every year numerous protein engineering and directed evolution studies are published, increasing the knowledge which could be used by protein engineers. Here we test a protein engineering strategy that allows quick access to improved biocatalysts with very little screening effort. Conceptually it is assumed that engineered residues, previously identified by rational and random methods, induce similar improvements when transferred to family members. Applied to ene-reductases from the Old Yellow Enzyme family (OYE), the newly created variants were tested with three compounds revealing more stereocomplementary OYE pairs with potent turnover frequencies (up to 660 h-1) and excellent stereoselectivities (up to >99%)...
January 20, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28107510/effect-of-the-crystal-environment-on-side-chain-conformational-dynamics-in-cyanovirin-n-investigated-through-crystal-and-solution-molecular-dynamics-simulations
#12
Logan S Ahlstrom, Ivan I Vorontsov, Jun Shi, Osamu Miyashita
Side chains in protein crystal structures are essential for understanding biochemical processes such as catalysis and molecular recognition. However, crystal packing could influence side-chain conformation and dynamics, thus complicating functional interpretations of available experimental structures. Here we investigate the effect of crystal packing on side-chain conformational dynamics with crystal and solution molecular dynamics simulations using Cyanovirin-N as a model system. Side-chain ensembles for solvent-exposed residues obtained from simulation largely reflect the conformations observed in the X-ray structure...
2017: PloS One
https://www.readbyqxmd.com/read/28106824/identification-of-the-muc2-promoter-as-a-strong-promoter-for-intestinal-gene-expression-through-generation-of-transgenic-quail-expressing-gfp-in-gut-epithelial-cells
#13
Rachel M Woodfint, Paula R Chen, Jinsoo Ahn, Yeunsu Suh, Seongsoo Hwang, Sang Suk Lee, Kichoon Lee
Identification of tissue- and stage-specific gene promoters is valuable for delineating the functional roles of specific genes in genetically engineered animals. Here, through the comparison of gene expression in different tissues by analysis of a microarray database, the intestinal specificity of mucin 2 (MUC2) expression was identified in mice and humans, and further confirmed in chickens by RT-PCR (reverse transcription-PCR) analysis. An analysis of cis-acting elements in avian MUC2 gene promoters revealed conservation of binding sites, within a 2...
January 19, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28106821/a-protective-vaccine-against-chlamydia-genital-infection-using-vault-nanoparticles-without-an-added-adjuvant
#14
Janina Jiang, Guangchao Liu, Valerie A Kickhoefer, Leonard H Rome, Lin-Xi Li, Stephen J McSorley, Kathleen A Kelly
Chlamydia trachomatis genital infection is the most common sexually transmitted bacterial disease, causing a significant burden to females due to reproductive dysfunction. Intensive screening and antibiotic treatment are unable to completely prevent female reproductive dysfunction, thus, efforts have become focused on developing a vaccine. A major impediment is identifying a safe and effective adjuvant which induces cluster of differentiation 4 (CD4) cells with attributes capable of halting genital infection and inflammation...
January 19, 2017: Vaccines
https://www.readbyqxmd.com/read/28106122/comparative-genomics-and-transcriptome-analysis-of-aspergillus-niger-and-metabolic-engineering-for-citrate-production
#15
Xian Yin, Hyun-Dong Shin, Jianghua Li, Guocheng Du, Long Liu, Jian Chen
Despite a long and successful history of citrate production in Aspergillus niger, the molecular mechanism of citrate accumulation is only partially understood. In this study, we used comparative genomics and transcriptome analysis of citrate-producing strains-namely, A. niger H915-1 (citrate titer: 157 g L(-1)), A1 (117 g L(-1)), and L2 (76 g L(-1))-to gain a genome-wide view of the mechanism of citrate accumulation. Compared with A. niger A1 and L2, A. niger H915-1 contained 92 mutated genes, including a succinate-semialdehyde dehydrogenase in the γ-aminobutyric acid shunt pathway and an aconitase family protein involved in citrate synthesis...
January 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28105787/orthogonal-surface-tags-for-whole-cell-biocatalysis
#16
Theo Peschke, Kersten S Rabe, Christof M Niemeyer
We herein describe the engineering of E. coli strains that display orthogonal tags for immobilization on their surface and overexpress a functional heterologous "protein content" in their cytosol at the same time. Using the outer membrane protein Lpp-ompA, cell-surface display of the streptavidin-binding peptide, the SpyTag/SpyCatcher system, or a HaloTag variant allowed us to generate bacterial strains that can selectively bind to solid substrates, as demonstrated with magnetic microbeads. The simultaneous cytosolic expression of functional content was demonstrated for fluorescent proteins or stereoselective ketoreductase enzymes...
January 20, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28103571/identification-of-aurintricarboxylic-acid-as-a-selective-inhibitor-of-the-tweak-fn14-signaling-pathway-in-glioblastoma-cells
#17
Alison Roos, Harshil D Dhruv, Ian T Mathews, Landon J Inge, Serdar Tuncali, Lauren K Hartman, Donald Chow, Nghia Millard, Holly H Yin, Jean Kloss, Joseph C Loftus, Jeffrey A Winkles, Michael E Berens, Nhan L Tran
The survival of patients diagnosed with glioblastoma (GBM), the most deadly form of brain cancer, is compromised by the proclivity for local invasion into the surrounding normal brain, which prevents complete surgical resection and contributes to therapeutic resistance. Tumor necrosis factor-like weak inducer of apoptosis (TWEAK), a member of the tumor necrosis factor (TNF) superfamily, can stimulate glioma cell invasion and survival via binding to fibroblast growth factor-inducible 14 (Fn14) and subsequent activation of the transcription factor NF-κB...
January 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/28103251/active-hydrogen-bond-network-ahbn-and-applications-for-improvement-of-thermal-stability-and-ph-sensitivity-of-pullulanase-from-bacillus-naganoensis
#18
Qing-Yan Wang, Neng-Zhong Xie, Qi-Shi Du, Yan Qin, Jian-Xiu Li, Jian-Zong Meng, Ri-Bo Huang
A method, so called "active hydrogen bond network" (AHBN), is proposed for site-directed mutations of hydrolytic enzymes. In an enzyme the AHBN consists of the active residues, functional residues, and conservative water molecules, which are connected by hydrogen bonds, forming a three dimensional network. In the catalysis hydrolytic reactions of hydrolytic enzymes AHBN is responsible for the transportation of protons and water molecules, and maintaining the active and dynamic structures of enzymes. The AHBN of pullulanase BNPulA324 from Bacillus naganoensis was constructed based on a homologous model structure using Swiss Model Protein-modeling Server according to the template structure of pullulanase BAPulA (2WAN)...
2017: PloS One
https://www.readbyqxmd.com/read/28102754/enhancement-of-antibody-functions-through-fc-multiplications
#19
Qun Wang, Yan Chen, Mark Pelletier, Romana Cvitkovic, Jessica Bonnell, Chien-Ying Chang, Adem C Koksal, Ellen O'Connor, Xizhe Gao, Xiang-Qing Yu, Herren Wu, C Kendall Stover, William F Dall'Acqua, Xiaodong Xiao
Antibodies carry out a plethora of functions through their crystallizable fragment (Fc) regions, which can be naturally tuned by the adoption of several isotypes and post-translational modifications. Protein engineering enables further Fc function modulations through modifications of the interactions between the Fc and its functional partners, including FcγR, FcRn, complement complex, and additions of auxiliary functional units. Due to the many functions embedded within the confinement of an Fc, a suitable balance must be maintained for a therapeutic antibody to be effective and safe...
January 19, 2017: MAbs
https://www.readbyqxmd.com/read/28102712/bone-morphogenetic-protein-2-and-7-mediate-the-anabolic-function-of-nucleus-pulposus-cells-with-discrete-mechanisms
#20
Victor Yl Leung, Lixiong Zhou, Wai-Kit Tam, Yi Sun, Fengjuan Lv, Guangqian Zhou, Kenneth Mc Cheung
Bone morphogenetic proteins (BMPs) play roles in promoting cell anabolism, especially extracellular matrix production. The difference between BMP members in their capacity to modulate intervertebral disc cell activity is yet to be defined. BMP-7/OP-1 has been shown to retard disc degeneration. We compared the activity of BMP-7 with BMP-2 on nucleus pulposus (NP) cell phenotype and function, and investigated how they differentially affect the gene expression profiles of signaling cascade components in human NP cells under degenerative states...
January 19, 2017: Connective Tissue Research
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