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https://www.readbyqxmd.com/read/28646659/analysis-of-recombinant-monoclonal-antibodies-in-hydrophilic-interaction-chromatography-a-generic-method-development-approach
#1
Balázs Bobály, Valentina D'Atri, Alain Beck, Davy Guillarme, Szabolcs Fekete
Hydrophilic interaction liquid chromatography (HILIC) is a well-established technique for the separation and analysis of small polar compounds. A recently introduced widepore stationary phase expanded HILIC applications to larger molecules, such as therapeutic proteins. In this paper, we present some generic HILIC conditions adapted for a wide range of FDA and EMA approved recombinant monoclonal antibody (mAb) species and for an antibody-drug conjugate (ADC). Seven approved mAbs possessing various isoelectric point (pI) and hydrophobicity as well as a cysteine conjugated ADC were used in this study...
June 15, 2017: Journal of Pharmaceutical and Biomedical Analysis
https://www.readbyqxmd.com/read/28646653/cryo-em-beyond-the-microscope
#2
REVIEW
Lesley A Earl, Veronica Falconieri, Jacqueline Ls Milne, Sriram Subramaniam
The pace at which cryo-EM is being adopted as a mainstream tool in structural biology has continued unabated over the past year. Initial successes in obtaining near-atomic resolution structures with cryo-EM were enabled to a large extent by advances in microscope and detector technology. Here, we review some of the complementary technical improvements that are helping sustain the cryo-EM revolution. We highlight advances in image processing that permit high resolution structure determination even in the presence of structural and conformational heterogeneity...
June 21, 2017: Current Opinion in Structural Biology
https://www.readbyqxmd.com/read/28646508/hepatic-stimulator-substance-resists-hepatic-ischemia-reperfusion-injury-by-regulating-drp1-translocation-and-activation
#3
Chao Zhang, Jing Huang, Wei An
Ischemia-reperfusion injury (IRI), induced by abnormal mitochondrial fission related apoptosis, is a major concern in liver transplantation settings. Our previous studies have demonstrated that hepatic stimulator substance (HSS) is an anti-apoptotic effector and could protect liver from IRI. However, the underlying mechanism remains unclear. In the present study, we report that in vitro and in vivo HSS could regulate mitochondrial fission and hepatocyte apoptosis during liver IRI by orchestrating the translocation and activation of dynamin-related protein 1 (Drp1)...
June 23, 2017: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
https://www.readbyqxmd.com/read/28646470/apobec3b-and-il-6-form-a-positive-feedback-loop-in-hepatocellular-carcinoma-cells
#4
Shuran Li, Xueyang Bao, Duowei Wang, Linjun You, Xianjing Li, Hongbao Yang, Jinsong Bian, Yun Wang, Yong Yang
APOBEC3 protein families, a DNA cytidine deaminase, were up-regulated in multiple tumors. However, the relationship between Hepatocellular carcinoma (HCC) and APOBEC3B (A3B) remains unknown. It has been confirmed that interleukin-6 (IL-6) has significant impacts on oncogenesis of HCC. Here, we reported that the expression of IL-6 was substantially up-regulated by A3B in HepG2 cells. A3B induced IL-6 expression through relocating HuR to enhance the IL-6 mRNA stability. Further analysis indicated that IL-6 also increased the expression of A3B through JAK1/STAT3 signaling pathway, which formed a positive feedback to maintain the continuous expression of A3B and IL-6, and thereby promoted the prolonged non-resolving inflammation...
May 29, 2017: Science China. Life Sciences
https://www.readbyqxmd.com/read/28646324/expression-purification-and-characterization-of-gmz2-10c-a-complex-disulphide-bonded-fusion-protein-vaccine-candidate-against-the-asexual-and-sexual-life-stages-of-the-malaria-causing-plasmodium-falciparum-parasite
#5
Ulrik H Mistarz, Susheel K Singh, Tam T T N Nguyen, Will Roeffen, Fen Yang, Casper Lissau, Søren M Madsen, Astrid Vrang, Régis W Tiendrebeogo, Ikhlaq H Kana, Robert W Sauerwein, Michael Theisen, Kasper D Rand
PURPOSE: Production and characterization of a chimeric fusion protein (GMZ2'.10C) which combines epitopes of key malaria parasite antigens: glutamate-rich protein (GLURP), merozoite surface protein 3 (MSP3), and the highly disulphide bonded Pfs48/45 (10C). GMZ2'.10C is a potential candidate for a multi-stage malaria vaccine that targets both transmission and asexual life-cycle stages of the parasite. METHODS: GMZ2'.10C was produced in Lactococcus lactis and purified using either an immunoaffinity purification (IP) or a conventional purification (CP) method...
June 23, 2017: Pharmaceutical Research
https://www.readbyqxmd.com/read/28645735/serum-amyloid-a-self-assembles-with-phospholipids-to-form-stable-protein-rich-nanoparticles-with-a-distinct-structure-a-hypothetical-function-of-saa-as-a-molecular-mop-in-immune-response
#6
Nicholas M Frame, Shobini Jayaraman, Donald L Gantz, Olga Gursky
Serum amyloid A (SAA) is an acute-phase protein whose action in innate immunity and lipid homeostasis is unclear. Most circulating SAA binds plasma high-density lipoproteins (HDL) and reroutes lipid transport. In vivo SAA binds existing lipoproteins or generates them de novo upon lipid uptake from cells. We explored the products of SAA-lipid interactions and lipoprotein remodeling in vitro. SAA complexes with palmitoyl-oleoyl phosphocholine (POPC) were analyzed for structure and stability using circular dichroism and fluorescence spectroscopy, electron microscopy, gel electrophoresis and gel filtration...
June 20, 2017: Journal of Structural Biology
https://www.readbyqxmd.com/read/28645531/new-protein-structures-provide-an-updated-understanding-of-phenylketonuria
#7
REVIEW
Eileen K Jaffe
Phenylketonuria (PKU) and less severe hyperphenylalaninemia (HPA) constitute the most common inborn error of amino acid metabolism, and is most often caused by defects in phenylalanine hydroxylase (PAH) function resulting in accumulation of Phe to neurotoxic levels. Despite the success of dietary intervention in preventing permanent neurological damage, individuals living with PKU clamor for additional non-dietary therapies. The bulk of disease-associated mutations are PAH missense variants, which occur throughout the entire 452 amino acid human PAH protein...
June 15, 2017: Molecular Genetics and Metabolism
https://www.readbyqxmd.com/read/28645370/xenopus-laevis-as-model-system-to-study-dna-damage-response-and-replication-fork-stability
#8
Vincenzo Sannino, Federica Pezzimenti, Stefania Bertora, Vincenzo Costanzo
Although many players of the DNA damage response and DNA repair have been identified in several systems their biochemical role is still poorly understood. The use of the Xenopus laevis egg extract cell-free system allowed biochemical dissection of DNA replication and cell cycle events in a complex biological context. The possibility of manipulating the protein content by using protein depletion procedures makes egg extract a powerful system to study proteins whose inactivation results in cellular lethality...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28645333/sac-1004-a-vascular-leakage-blocker-reduces-cerebral-ischemia-reperfusion-injury-by-suppressing-blood-brain-barrier-disruption-and-inflammation
#9
Haiying Zhang, Joon Ha Park, Sony Maharjan, Jeong Ae Park, Kyu-Sung Choi, Hyojin Park, Yoonjeong Jeong, Ji Hyeon Ahn, In Hye Kim, Jae-Chul Lee, Jeong Hwi Cho, In-Kyu Lee, Choong Hyun Lee, In Koo Hwang, Young-Myeong Kim, Young-Ger Suh, Moo-Ho Won, Young-Guen Kwon
BACKGROUND: Blood-brain barrier (BBB) breakdown and inflammation are critical events in ischemic stroke, contributing to aggravated brain damage. The BBB mainly consists of microvascular endothelial cells sealed by tight junctions to protect the brain from blood-borne substances. Thus, the maintenance of BBB integrity may be a potential target for neuroprotection. Sac-1004, a pseudo-sugar derivative of cholesterol, enhances the endothelial barrier by the stabilization of the cortical actin ring...
June 23, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28645217/microtubule-organizing-centers
#10
Jingchao Wu, Anna Akhmanova
The organization of microtubule networks is crucial for controlling chromosome segregation during cell division, positioning, and transport of different organelles and for cell polarity and morphogenesis. The geometry of microtubule arrays strongly depends on the localization and activity of the sites where microtubules are nucleated and where their minus ends are anchored. Such sites are often clustered into structures known as microtubule-organizing centers, which include the centrosomes in animals and spindle pole bodies in fungi...
June 23, 2017: Annual Review of Cell and Developmental Biology
https://www.readbyqxmd.com/read/28645112/tap63%C3%AE-and-%C3%AE-np63%C3%AE-are-regulated-by-rbm38-via-mrna-stability-and-have-an-opposing-function-in-growth-suppression
#11
Wensheng Yan, Yanhong Zhang, Xinbin Chen
The p63 gene is expressed as TAp63 from the P1 promoter and as ΔNp63 from the P2 promoter. Through alternative splicing, five TA and five ΔN isoforms (α-ε) are expressed. Isoforms α-β and δ share an identical 3' untranslated region (3'UTR) whereas isoform γ has a unique 3'UTR. Recently, we found that RBM38 RNA-binding protein is a target of p63 and RBM38 in turn regulates p63α/β expression via mRNA stability. However, it is uncertain whether p63γ has a unique biological activity and whether p63γ is regulated by RBM38...
June 13, 2017: Oncotarget
https://www.readbyqxmd.com/read/28644968/serum-lipoprotein-associated-phospholipase-a2-predicts-the-formation-of-carotid-artery-plaque-and-its-vulnerability-in-anterior-circulation-cerebral-infarction
#12
Yuping Yang, Tao Xue, Juehua Zhu, Jiayi Xu, Xiaowei Hu, Penghao Wang, Tao Kong, Yan Yan, Lihui Yang, Shouru Xue
OBJECTIVE: Circulation inflammation markers such as high-sensitive C-reactive protein (hsCRP) and lipoprotein-associated phospholipase A2 (Lp-PLA2) are considered as predictors of cerebral and cardiac vascular diseases. However, the role of hsCRP and Lp-PLA2 in the anterior circulation cerebral infarction (ACI) is to be elaborated. PATIENTS AND METHODS: We included 100 patients with acute anterior circulation cerebral infarction (AaCI group) and 50 non-infarction subjects (control group)...
June 12, 2017: Clinical Neurology and Neurosurgery
https://www.readbyqxmd.com/read/28644630/expanding-the-therapeutic-potential-of-the-iron-chelator-deferasirox-in-the-development-of-aqueous-stable-ti-iv-anticancer-complexes
#13
Sergio A Loza-Rosas, Alexandra M Vázquez-Salgado, Kennett I Rivero, Lenny J Negrón, Yamixa Delgado, Josué A Benjamín-Rivera, Angel L Vázquez-Maldonado, Timothy B Parks, Charlene Munet-Colón, Arthur D Tinoco
The recent X-ray structure of titanium(IV)-bound human serum transferrin (STf) exhibiting citrate as a synergistic anion reveals a difference in Ti(IV) coordination versus iron(III), the metal endogenously delivered by the protein to cells. This finding enriches our bioinspired drug design strategy for Ti(IV)-based anticancer therapeutics, which applies a family of Fe(III) chelators termed chemical transferrin mimetic (cTfm) ligands to inhibit Fe bioavailability in cancer cells. Deferasirox, a drug used for iron overload disease, is a cTfm ligand that models STf coordination to Fe(III), favoring Fe(III) binding versus Ti(IV)...
June 23, 2017: Inorganic Chemistry
https://www.readbyqxmd.com/read/28644385/ch-%C3%AF-interactions-in-carbohydrate-recognition
#14
REVIEW
Vojtěch Spiwok
Many carbohydrate-binding proteins contain aromatic amino acid residues in their binding sites. These residues interact with carbohydrates in a stacking geometry via CH/π interactions. These interactions can be found in carbohydrate-binding proteins, including lectins, enzymes and carbohydrate transporters. Besides this, many non-protein aromatic molecules (natural as well as artificial) can bind saccharides using these interactions. Recent computational and experimental studies have shown that carbohydrate-aromatic CH/π interactions are dispersion interactions, tuned by electrostatics and partially stabilized by a hydrophobic effect in solvated systems...
June 23, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28644022/hot-spot-residues-to-be-mutated-common-in-g-protein-coupled-receptors-of-class-a-identification-of-thermostabilizing-mutations-followed-by-determination-of-three-dimensional-structures-for-two-example-receptors
#15
Satoshi Yasuda, Yuta Kajiwara, Yosuke Toyoda, Kazushi Morimoto, Ryoji Suno, So Iwata, Takuya Kobayashi, Takeshi Murata, Masahiro Kinoshita
G protein-coupled receptors (GPCRs), which are indispensable to life and also implicated in a number of diseases, construct important drug targets. For the efficient structure-guided drug design, however, their structural stabilities must be enhanced. An amino-acid mutation is known to possibly lead to the enhancement, but currently available experimental and theoretical methods for identifying stabilizing mutations suffer such drawbacks as the incapability of exploring the whole mutational space with minor effort and the unambiguous physical origin of the enhanced or lowered stability...
June 23, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28644007/cyclotides-as-tools-in-chemical-biology
#16
Simon J de Veer, Joachim Weidmann, David J Craik
Among the various molecules that plants produce for defense against pests and pathogens, cyclotides stand out as exceptionally stable and structurally unique. These ribosomally synthesized peptides are around 30 amino acids in size, and are stabilized by a head-to-tail cyclic peptide backbone and three disulfide bonds that form a cystine knot. They occur in certain plants of the Rubiaceae, Violaceae, Cucurbitaceae, Fabaceae, and Solanaceae families, with an individual plant producing up to hundreds of different cyclotides...
June 23, 2017: Accounts of Chemical Research
https://www.readbyqxmd.com/read/28643783/cdt1-stabilizes-an-open-mcm-ring-for-helicase-loading
#17
Jordi Frigola, Jun He, Kerstin Kinkelin, Valerie E Pye, Ludovic Renault, Max E Douglas, Dirk Remus, Peter Cherepanov, Alessandro Costa, John F X Diffley
ORC, Cdc6 and Cdt1 act together to load hexameric MCM, the motor of the eukaryotic replicative helicase, into double hexamers at replication origins. Here we show that Cdt1 interacts with MCM subunits Mcm2, 4 and 6, which both destabilizes the Mcm2-5 interface and inhibits MCM ATPase activity. Using X-ray crystallography, we show that Cdt1 contains two winged-helix domains in the C-terminal half of the protein and a catalytically inactive dioxygenase-related N-terminal domain, which is important for MCM loading, but not for subsequent replication...
June 23, 2017: Nature Communications
https://www.readbyqxmd.com/read/28643740/telomere-length-variation-and-expression-analysis-of-shelterin-complex-genes-during-gallbladder-carcinogenesis
#18
Satish S Poojary, Gunja Mishra, Tekcham Dinesh Singh, Sanjiv Gupta, Braj Raj Shrivastav, Pramod Kumar Tiwari
BACKGROUND: Telomeres, which are bound with shelterin protein complex, play an important role in maintaining genomic stability and its dysfunction may lead to carcinogenesis. Here, we aimed to analyze whether shelterin complex gene expression and telomere length variation, play any role in gallbladder carcinogenesis. METHODS: Telomere length analysis was carried out by monochrome multiplex qPCR, whereas expression analysis of shelterin genes was carried out using RT-qPCR...
April 2017: Journal of Cancer Research and Therapeutics
https://www.readbyqxmd.com/read/28643574/stability-and-in-vitro-release-behaviour-of-encapsulated-omega-fatty-acid-rich-oils-in-lentil-protein-isolate-based-microcapsules
#19
C Chang, M T Nickerson
The objective of this study was to investigate the use of a lentil protein isolate-based microcapsule design as a platform for entrapping different types of omega fatty acid-rich oil (e.g. canola, fish and flaxseed oils) and to characterise differences in the physical properties (e.g. moisture content, water activity, colour, wettability, particle size, surface oil and entrapment efficiency), storage stability and in vitro release behaviour of the entrapped oils. All microcapsules displayed similar physical properties regardless of the core material...
June 23, 2017: International Journal of Food Sciences and Nutrition
https://www.readbyqxmd.com/read/28643528/modulating-release-of-ranibizumab-and-aflibercept-from-thiolated-chitosan-based-hydrogels-for-potential-treatment-of-ocular-neovascularization
#20
Miguel Moreno, Poh Yih Pow, Tan Su Teng Tabitha, Sonali Nirmal, Andreas Larsson, Krishna Radhakrishnan, Jayabalan Nirmal, Soo Tng Quah, Susana Geifman Shochat, Rupesh Agrawal, Subbu Venkatraman
BACKGROUND: This paper describes the synthesis of thiolated chitosan-based hydrogels with varying degrees of crosslinking that has been utilized to modulate release kinetics of two clinically relevant FDA-approved anti-VEGF protein drugs, ranibizumab and aflibercept. These hydrogels have been fabricated into disc shaped structures for potential use as patches on ocular surface. METHODS: Protein conformational changes and aggregation after loading and release was evaluated by circular dichroism (CD), steady-state tryptophan fluorescence spectroscopy, electrophoresis and size-exclusion chromatography (SEC)...
June 23, 2017: Expert Opinion on Drug Delivery
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