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Protein Journal

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https://www.readbyqxmd.com/read/29139018/vaccination-against-hydatidosis-molecular-cloning-and-optimal-expression-of-the-eg95nc-recombinant-antigen-in-escherichia-coli
#1
M Jazouli, M Lightowlers, C G Gauci, K Tadlaoui, A Belmlih, M M Ennaji, M Elharrak
Cystic echinococcosis (CE) is a widely distributed zoonosis that is highly endemic in the Mediterranean basin. The disease represents a serious public health threat and causes economic losses. The parasite life-cycle involves dogs and ruminants as definitive and intermediate hosts; humans are accidently infected, causing serious clinical issues. Vaccination of ruminants and dog treatments represent the most efficient measures to prevent parasite transmission. The recombinant protein vaccine, EG95, has been used successfully in sheep vaccine trials against CE in several countries...
November 14, 2017: Protein Journal
https://www.readbyqxmd.com/read/29128960/a-proteomics-analysis-reveals-9-up-regulated-proteins-associated-with-altered-cell-signaling-in-colon-cancer-patients
#2
Oleg I Kit, Dmitry I Vodolazhsky, Denis S Kutilin, Yaroslav S Enin, Yury A Gevorkyan, Peter V Zolotukhin, Yanis Boumber, Leonid V Kharin, Svetlana B Panina
Colorectal cancer is the second most common cancer in women and third most common cancer in men. Cell signaling alterations in colon cancer, especially in aggressive metastatic tumors, require further investigations. The present study aims to compare the expression pattern of proteins associated with cell signaling in paired tumor and non-tumor samples of patients with colon cancer, as well as to define the cluster of proteins to differentiate patients with non-metastatic (Dukes' grade B) and metastatic (Dukes' grade C&D) colon cancer...
November 11, 2017: Protein Journal
https://www.readbyqxmd.com/read/29127553/identification-of-histidine-303-as-the-catalytic-base-of-lysyl-oxidase-via-site-directed-mutagenesis
#3
Rachel N Oldfield, Kathryn A Johnston, Jeanette Limones, Caitlin Ghilarducci, Karlo M Lopez
Lysyl oxidase (LOX) is a copper-dependent amine oxidase enzyme that catalyzes the formation of crosslinkages of collagen and elastin in connective tissues by oxidative deamination of lysine. Using site-directed mutagenesis, Histidine 303 has been shown to be a key residue that acts as the necessary catalytic base for this enzyme to function properly. Histidine 303 was mutated to isoleucine to remove catalytic activity and to aspartate and glutamate, respectively, in order to provide alternate residues that could act as a general base that could maintain catalytic activity...
November 10, 2017: Protein Journal
https://www.readbyqxmd.com/read/29119487/exploring-the-roles-of-proline-in-three-dimensional-domain-swapping-from-structure-analysis-and-molecular-dynamics-simulations
#4
Yongqi Huang, Meng Gao, Zhengding Su
Three-dimensional (3D) domain swapping is a mechanism to form protein oligomers. It has been proposed that several factors, including proline residues in the hinge region, may affect the occurrence of 3D domain swapping. Although introducing prolines into the hinge region has been found to promote domain swapping for some proteins, the opposite effect has also been observed in several studies. So far, how proline affects 3D domain swapping remains elusive. In this work, based on a large set of 3D domain-swapped structures, we performed a systematic analysis to explore the correlation between the presence of proline in the hinge region and the occurrence of 3D domain swapping...
November 8, 2017: Protein Journal
https://www.readbyqxmd.com/read/29052020/erratum-to-a-rapid-and-reliable-method-for-total-protein-extraction-from-succulent-plants-for-proteomic-analysis
#5
Fernando Lledías, Felipe Hernández, Viridiana Rivas, Abisaí García-Mendoza, Gladys I Cassab, Jorge Nieto-Sotelo
The original version of this article unfortunately contains a mistake. The authors have inadvertently incorrectly listed the concentration of TCA in the acetone/TCA/β-ME solution in the materials and methods section of this paper. The TCA concentration in Sects. 2.3.2 and 2.3.5 should be 10% TCA, making the solution acetone/10% TCA/0.07% β-ME. It is now corrected with this erratum.
October 19, 2017: Protein Journal
https://www.readbyqxmd.com/read/29030733/high-yield-preparation-of-functionally-active-catalytic-translocation-domain-module-of-botulinum-neurotoxin-type-a-that-exhibits-uniquely-different-enzyme-kinetics
#6
Harkiranpreet Kaur Dhaliwal, Nagarajan Thiruvanakarasu, Raj Kumar, Kruti Patel, Ghuncha Ambrin, Shouwei Cai, Bal Ram Singh
Botulinum neurotoxins (BoNTs) are the most toxic proteins known to cause flaccid muscle paralysis as a result of inhibition of neurotransmitter release from peripheral cholinergic synapses. BoNT type A (BoNT/A) is a 150 kDa protein consisting of two major subunits: light chain (LC) and heavy chain (HC). The LC is required for the catalytic activity of neurotoxin, whereas the C and N terminal domains of the HC are required for cell binding, and translocation of LC across the endosome membranes, respectively...
October 13, 2017: Protein Journal
https://www.readbyqxmd.com/read/29022133/arginine-kinases-from-the-precious-corals-corallium-rubrum-and-paracorallium-japonicum-presence-of-two-distinct-arginine-kinase-gene-lineages-in-cnidarians
#7
Tomoka Matsuo, Daichi Yano, Kouji Uda, Nozomu Iwasaki, Tomohiko Suzuki
The cDNA sequence of arginine kinase (AK) from the precious coral Corallium rubrum was assembled from transcriptome sequence data, and the deduced amino acid sequence of 364 residues was shown to conserve the structural features characteristic of AK. Based on the amino acid sequence, the DNA coding C. rubrum AK was synthesized by overlap extension PCR to prepare the recombinant enzyme. The following kinetic parameters were determined for the C. rubrum enzyme: K a(Arg) (0.10 mM), K ia(Arg) (0.79 mM), K a(ATP) (0...
October 11, 2017: Protein Journal
https://www.readbyqxmd.com/read/28975545/heterologous-expression-purification-and-characterization-of-a-peroxidase-isolated-from-lepidium-draba
#8
Yaser Fattahian, Ali Riahi-Madvar, Reza Mirzaee, Masoud Torkzadeh-Mahani, Gholamreza Asadikaram
Peroxidase is one of the most widely used enzymes in biotechnology and medicine. In the current study, cDNA encoding peroxidase from Lepidium draba (LDP) was cloned and expressed in Escherichia coli BL21 (DE3) cells in the form of inclusion bodies (IBs). To achieve purified active enzyme, IBs were solubilized before being purified and refolded. The deduced amino acid sequence (308) of the LDP gene (924 bp) revealed 88.96% identity to horseradish peroxidase C1A (HRP C1A). The results of basic local alignment search tool (BLAST) and phylogenetic analysis of the protein sequence showed that this enzyme belongs to the neutral group of class III plant peroxidases...
October 3, 2017: Protein Journal
https://www.readbyqxmd.com/read/28975457/characterization-of-a-novel-alkalophilic-lipase-from-aneurinibacillus-thermoaerophilus-lid-heterogeneity-and-assignment-to-family-i-5
#9
Ximena Zottig, Fatma Meddeb-Mouelhi, David M Charbonneau, Marc Beauregard
Recent investigations of Aneurinibacillus thermoaerophilus strains have allowed identification of a unique solvent tolerant lipase, distinct from known lipases. This work reports the expression and purification of this lipase (LipAT) and the first characterization of its structure and temperature and pH-dependent behaviour. LipAT has a secondary structural content compatible with the canonical lipase α/β hydrolase fold, and is dimeric at neutral pH. The protein was folded from pH 5 to 10, and association into folded aggregates at pH 7 and 8 likely protected its secondary structures from thermal unfolding...
October 3, 2017: Protein Journal
https://www.readbyqxmd.com/read/28932939/the-thumb-domain-is-not-essential-for-the-catalytic-action-of-holama-dna-polymerase
#10
Angela Gala Morena Gatius, Fabrizio Dal Piaz, Alejandro Hochkoeppler
A structural and kinetic characterization of a fragment of the HoLaMa DNA polymerase is presented here. In particular, a truncated form of HoLaMa, devoid of a consistent portion of the thumb domain, was isolated and purified. This HoLaMa fragment, denoted as ΔNter-HoLaMa, is surprisingly competent in catalyzing DNA extension, albeit featuring a kcat one order of magnitude lower than the corresponding kinetic constant of its full-length counterpart. The conformational rearrangements, if any, of enzyme tryptophanes triggered by DNA binding or extension were assayed under pre-steady-state conditions...
September 20, 2017: Protein Journal
https://www.readbyqxmd.com/read/28856545/oxidative-folding-of-conopeptides-modified-by-conus-protein-disulfide-isomerase
#11
Lei Wang, Xiaomin Wang, Zhenghua Ren, Wei Tang, Qiong Zou, Jinxing Wang, Shangwu Chen, Han Zhang, Anlong Xu
Protein disulfide isomerase is a type of enzyme that catalyses the oxidation, isomerization and reduction of disulfide bonds. Conotoxins that containing disulfide bonds are likely substrates of protein disulfide isomerise. Here, we cloned 12 protein disulfide isomerise genes from 12 different cone snail species that inhabited the sea near Sanya in China. The full-length amino acid sequences of these protein disulfide isomerase genes share a high degree of homology, including the same -CGHC- active site sequence and -RDEL- endoplasmic reticulum retention signal...
October 2017: Protein Journal
https://www.readbyqxmd.com/read/28852914/induction-of-metallothionein-in-rat-liver-by-zinc-exposure-a-dose-and-time-dependent-study
#12
Roobee Garla, Preeti Kango, Navneet Kaur Gill, M L Garg
Metallothioneins (MTs) are low molecular weight ubiquitous metalloproteins with high cysteine (thiol) content. The intracellular concentration of zinc (Zn) is tightly regulated and MT plays a crucial role in it. The present study investigates the relationship between the Zn status (as a function of Zn concentration and time) in the rat liver and the occurrence of hepatic MT. For dose dependent study, four experimental groups, one control and three receiving different levels of metal supplementation, were chosen [Group 1 control and Group 2, Group 3, Group 4 receiving subcutaneous dose of 10, 50 and 100 mg of Zn/kg body weight (in the form of ZnSO4·7H2O), respectively]...
October 2017: Protein Journal
https://www.readbyqxmd.com/read/28815420/road-map-for-the-structure-based-design-of-selective-covalent-hcv-ns3-4a-protease-inhibitors
#13
REVIEW
Letitia Shunmugam, Pritika Ramharack, Mahmoud E S Soliman
Over the last 2 decades, covalent inhibitors have gained much popularity and is living up to its reputation as a powerful tool in drug discovery. Covalent inhibitors possess many significant advantages including increased biochemical efficiency, prolonged duration and the ability to target shallow, solvent exposed substrate-binding domains. However, rapidly mounting concerns over the potential toxicity, highly reactive nature and general lack of selectivity have negatively impacted covalent inhibitor development...
October 2017: Protein Journal
https://www.readbyqxmd.com/read/28762197/sequence-structural-analysis-and-metrics-to-define-the-unique-dynamic-features-of-the-flap-regions-among-aspartic-proteases
#14
REVIEW
Lara McGillewie, Muthusamy Ramesh, Mahmoud E Soliman
Aspartic proteases are a class of hydrolytic enzymes that have been implicated in a number of diseases such as HIV, malaria, cancer and Alzheimer's. The flap region of aspartic proteases is a characteristic unique structural feature of these enzymes; and found to have a profound impact on protein overall structure, function and dynamics. Flap dynamics also plays a crucial role in drug binding and drug resistance. Therefore, understanding the structure and dynamic behavior of this flap regions is crucial in the design of potent and selective inhibitors against aspartic proteases...
October 2017: Protein Journal
https://www.readbyqxmd.com/read/28755312/mouse-nudt13-is-a-mitochondrial-nudix-hydrolase-with-nad-p-h-pyrophosphohydrolase-activity
#15
Salama R Abdelraheim, David G Spiller, Alexander G McLennan
The mammalian NUDT13 protein possesses a sequence motif characteristic of the NADH pyrophosphohydrolase subfamily of Nudix hydrolases. Due to the persistent insolubility of the recombinant product expressed in Escherichia coli, active mouse Nudt13 was expressed in insect cells from a baculovirus vector as a histidine-tagged recombinant protein. In vitro, it efficiently hydrolysed NADH to NMNH and AMP and NADPH to NMNH and 2',5'-ADP and had a marked preference for the reduced pyridine nucleotides. Much lower activity was obtained with other nucleotide substrates tested...
October 2017: Protein Journal
https://www.readbyqxmd.com/read/28710679/new-phage-display-isolated-heptapeptide-recognizing-the-regulatory-carboxy-terminal-domain-of-human-tumour-protein-p53
#16
Sihem Ben Abid, Mouna Sahnoun, Ines Yacoubi-Hadj Amor, Salma Abdelmoula-Souissi, Hajer Hassairi, Raja Mokdad-Gargouri, Ali Gargouri
The transcription factor tumor protein p53 (P53) controls a variety of genes most involved in cell cycle and is at the origin of apoptosis when DNA is irreparably damaged. We planned to select novel tumor protein p53-interacting peptides through the screening of hepta-peptide phage-display libraries. For this aim, human tumor suppressor protein p53 was expressed in Escherichia coli as Glutathione S-transferase fusion and purified by affinity chromatography. The phage library was then screened on this immobilized protein target...
October 2017: Protein Journal
https://www.readbyqxmd.com/read/28707196/reversible-association-of-the-hemagglutinin-subcomplex-ha-33-ha-17-trimer-with-the-botulinum-toxin-complex
#17
Yoshimasa Sagane, Shingo Mutoh, Ryosuke Koizumi, Tomonori Suzuki, Shin-Ichiro Miyashita, Keita Miyata, Tohru Ohyama, Koichi Niwa, Toshihiro Watanabe
Botulinum neurotoxin (BoNT) associates with nontoxic proteins, either a nontoxic nonhemagglutinin (NTNHA) or the complex of NTNHA and hemagglutinin (HA), to form M- or L-toxin complexes (TCs). Single BoNT and NTNHA molecules are associated and form M-TC. A trimer of the 70-kDa HA protein (HA-70) attaches to the M-TC to form M-TC/HA-70. Further, 1-3 arm-like 33- and 17-kDa HA molecules (HA-33/HA-17 trimer), consisting of 1 HA-17 protein and 2 HA-33 proteins, can attach to the M-TC/HA-70 complex, yielding 1-, 2-, and 3-arm L-TC...
October 2017: Protein Journal
https://www.readbyqxmd.com/read/28699078/characterization-of-akr4c15-a-novel-member-of-aldo-keto-reductase-in-comparison-with-other-rice-akr-s
#18
COMPARATIVE STUDY
Budsakorn Auiyawong, Rawint Narawongsanont, Chonticha Tantitadapitak
Environmental stresses often cause a rapid and excessive accumulation of reactive oxygen species (ROS), the toxicity of which is further amplified by downstream aldehyde production. Aldo-keto reductase (AKR) is a group of enzymes metabolizing aldehyde/ketone to the corresponding alcohol using NADPH as the cofactor. In this study, OsI_20197 (AKR4C15), a novel member of AKR4 subfamily C, was isolated and biochemically characterized. Kinetic studies on bacterially-expressed recombinant AKR4C15 revealed that the enzyme was capable of metabolizing a wide variety of aldehydes but clearly exhibited a preference for three carbon compounds, i...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28669025/effects-of-gsh1-and-gsh2-gene-mutation-on-glutathione-synthetases-activity-of-saccharomyces-cerevisiae
#19
Wen Xu, Haiyan Jia, Longmei Zhang, Haiyan Wang, Hui Tang, Liping Zhang
In this paper, three mutants from wild Saccharomyces cerevisiae HBU2.558, called U2.558, UN2.558, and UNA2.558, were screened by UV, sodium nitrite, Atmospheric and room temperature plasma, respectively. Glutathione production of the three mutants increased by 41.86, 72.09 and 56.76%, respectively. We detected the activity of glutathione synthetases and found that its activity was improved. Amino acid sequences of three mutant colonies were compared with HBU2.558. Four mutants: Leu51→Pro51 (L51P), Glu62→Val62 (E62V), Ala332→Glu332 (A332E) and Ser653→Gly653 (S653G) were found in the analysis of γ-glutamylcysteine ligase...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28660316/expression-of-flp-protein-in-a-baculovirus-insect-cell-system-for-biotechnological-applications
#20
Ida S Jensen, Ken Inui, Srdja Drakulic, Sakthidasan Jayaprakash, Bjoern Sander, Monika M Golas
The Saccharomyces cerevisiae Flp protein is a site-specific recombinase that recognizes and binds to the Flp recognition target (FRT) site, a specific sequence comprised of at least two inverted repeats separated by a spacer. Binding of four monomers of Flp is required to mediate recombination between two FRT sites. Because of its site-specific cleavage characteristics, Flp has been established as a genome engineering tool. Amongst others, Flp is used to direct insertion of genes of interest into eukaryotic cells based on single and double FRT sites...
August 2017: Protein Journal
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