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Protein Journal

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https://www.readbyqxmd.com/read/28710679/new-phage-display-isolated-heptapeptide-recognizing-the-regulatory-carboxy-terminal-domain-of-human-tumour-protein-p53
#1
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...
July 14, 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
#2
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...
July 13, 2017: Protein Journal
https://www.readbyqxmd.com/read/28567566/letter-from-the-new-editor-in-chief
#3
EDITORIAL
Lawrence J Berliner
No abstract text is available yet for this article.
June 1, 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
#4
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
#5
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
#6
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
https://www.readbyqxmd.com/read/28660315/characterization-of-the-dihydroorotase-from-methanococcus-jannaschii
#7
Jacqueline Vitali, Aditya K Singh, Michael J Colaneri
The gene that codes for the putative dihydroorotase in the hyperthermophilic archaeon Methanococcus jannaschii was subcloned in pET-21a and expressed in Escherichia coli. A purification protocol was devised. The purity of the protein was evaluated by SDS-PAGE and the protein was confirmed by sequencing using LC-MS. The calculated molecular mass is 48104 Da. SEC-LS suggested that the protein is a monomer in solution. ICP-MS showed that there are two Zn ions per monomer. Kinetic analysis of the recombinant protein gave hyperbolic kinetics with Vmax = 12...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28646265/omega-3-and-omega-6-fatty-acids-act-as-inhibitors-of-the-matrix-metalloproteinase-2-and-matrix-metalloproteinase-9-activity
#8
Eleonora Nicolai, Federica Sinibaldi, Gianpaolo Sannino, Giuseppina Laganà, Francesco Basoli, Silvia Licoccia, Paola Cozza, Roberto Santucci, Maria Cristina Piro
Polyunsaturated fatty acids have been reported to play a protective role in a wide range of diseases characterized by an increased metalloproteinases (MMPs) activity. The recent finding that omega-3 and omega-6 fatty acids exert an anti-inflammatory effect in periodontal diseases has stimulated the present study, designed to determine whether such properties derive from a direct inhibitory action of these compounds on the activity of MMPs. To this issue, we investigated the effect exerted by omega-3 and omega-6 fatty acids on the activity of MMP-2 and MMP-9, two enzymes that actively participate to the destruction of the organic matrix of dentin following demineralization operated by bacteria acids...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28639160/expression-and-activity-analysis-of-fructosyltransferase-from-aspergillus-oryzae
#9
Lihong Guan, Liping Chen, Yongsen Chen, Nu Zhang, Yawei Han
The fructosyltransferase gene was isolated and cloned from Aspergillus oryzae. The gene was 1368 bp, which encoded a protein of 455 amino acids. To analyze the activity of the expressed fructosyltransferase, the pET32a-fructosyltransferase recombined plasmid was transformed into Escherichia coli BL21. The fructosyltransferase gene was successfully expressed by Isopropyl-β-d-thiogalactoside (IPTG) induction. The molecular weight of the expression protein was about 45 kDa. The optimal conditions of protein expression were 25 °C, 0...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28634775/hydrophilic-acylated-surface-protein-a-haspa-of-leishmania-donovani-expression-purification-and-biophysico-chemical-characterization
#10
Manoj Kumar, Kishu Ranjan, Vijay Singh, Chandramani Pathak, Anju Pappachan, Desh Deepak Singh
Hydrophilic acylated surface proteins (HASPs) are acidic surface proteins which get localized on the surface of Leishmania parasite during infective stages through a "non-classical" pathway. In this study, we report the heterologous expression and purification of Leishmania donovani HASPA (r-LdHASPA) in E. coli system and its partial characterization. The structural aspects of the purified protein were analyzed using CD spectroscopy and modeling studies which indicate that r-LdHASPA consists of random coils...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28634774/learning-from-synthetic-models-of-extracellular-matrix-differential-binding-of-wild-type-and-amyloidogenic-human-apolipoprotein-a-i-to-hydrogels-formed-from-molecules-having-charges-similar-to-those-found-in-natural-gags
#11
Silvana A Rosú, Leandro Toledo, Bruno F Urbano, Susana A Sanchez, Graciela C Calabrese, M Alejandra Tricerri
Among other components of the extracellular matrix (ECM), glycoproteins and glycosaminoglycans (GAGs) have been strongly associated to the retention or misfolding of different proteins inducing the formation of deposits in amyloid diseases. The composition of these molecules is highly diverse and a key issue seems to be the equilibrium between physiological and pathological events. In order to have a model in which the composition of the matrix could be finely controlled, we designed and synthesized crosslinked hydrophilic polymers, the so-called hydrogels varying the amounts of negative charges and hydroxyl groups that are prevalent in GAGs...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28589291/hydroxylation-of-human-type-iii-collagen-alpha-chain-by-recombinant-coexpression-with-a-viral-prolyl-4-hydroxylase-in-escherichia-coli
#12
Jingjing Shi, Xiaoxuan Ma, Yuan Gao, Daidi Fan, Chenhui Zhu, Yu Mi, Wenjiao Xue
High-level expression of recombinant collagen by genetic engineering is urgently required. Recombinant collagen is different from natural collagen in its hydroxyproline (Hyp) content and thermal stability. To obtain hydroxylated collagen for applications in biomedicine and biomaterials, the human collagen α1(III) chain was co-expressed with the viral prolyl 4-hydroxylase A085R in Escherichia coli. Unlike previous reports using human prolyl 4-hydroxylase, this study examined the hydroxylation of full-length human collagen α1(III) chain (COL3A1) by viral prolyl 4-hydroxylase...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28555319/proteomic-analysis-of-rice-seedlings-under-cold-stress
#13
Li Ji, Ping Zhou, Ya Zhu, Fang Liu, Rongbai Li, Yongfu Qiu
Low temperature can greatly restrict the growth and development of rice. The rice seedlings show growth retardation, lamina wrap, and part of blade even died under the condition of low temperature. In order to get more information about cold stress responses in rice, two dimensional electrophoresis and bioinformatics analysis of mass spectrometry were used to preliminary survey the cold tolerance of cold sensitive line 9311 and cold resistance variety Fujisaka 5 under cold stress. Two dimensional electrophoresis maps of 9311 and Fujisaka 5 were established under cold treatment...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28555318/conformational-characteristics-of-rice-hexokinase-oshxk7-as-a-moonlighting-protein-involved-in-sugar-signalling-and-metabolism
#14
Li Wang, Qing Dong, Qingdong Zhu, Niwen Tang, Shenghua Jia, Chao Xi, Heping Zhao, Shengcheng Han, Yingdian Wang
Hexokinase (HXK) as a moonlighting protein involves in glucose metabolism and signalling to regulate growth and development in plants. Therefore, the clarification for the structural properties of OsHXK7 (Oryza sativa Hexokinase 7) is essential to understand its role mechanism associated with the Glc signalling and metabolism. In this study, the structural characteristics of OsHXK7 (Oryza sativa Hexokinase 7) were identified. In the fluorescence spectrum, the Trp peak representing OsHXK7 binding to D-glucose (D-Glc) and 2-deoxyglucose (2-dG) showed an obvious blue shift...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28497409/a-rapid-and-reliable-method-for-total-protein-extraction-from-succulent-plants-for-proteomic-analysis
#15
Fernando Lledías, Felipe Hernández, Viridiana Rivas, Abisaí García-Mendoza, Gladys I Cassab, Jorge Nieto-Sotelo
Crassulacean acid metabolism plants have some morphological features, such as succulent and reduced leaves, thick cuticles, and sunken stomata that help them prevent excessive water loss and irradiation. As molecular constituents of these morphological adaptations to xeric environments, succulent plants produce a set of specific compounds such as complex polysaccharides, pigments, waxes, and terpenoids, to name a few, in addition to uncharacterized proteases. Since all these compounds interfere with the analysis of proteins by electrophoretic techniques, preparation of high quality samples from these sources represents a real challenge...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28470375/subtractive-protein-profiling-of-salmonella-typhimurium-biofilm-treated-with-dmso
#16
Mohd Fakharul Zaman Raja Yahya, Zazali Alias, Saiful Anuar Karsani
Salmonella typhimurium is an important biofilm-forming bacteria. It is known to be resistant to a wide range of antimicrobials. The present study was carried out to evaluate the effects of dimethyl sulfoxide (DMSO) against S. typhimurium biofilm and investigate whole-cell protein expression by biofilm cells following treatment with DMSO. Antibiofilm activities were assessed using pellicle assay, crystal violet assay, colony-forming unit counting and extracellular polymeric substance (EPS) matrix assay whilst differential protein expression was investigated using a combination of one dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, tandem mass spectrometry and bioinformatics...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28429156/crystallization-and-preliminary-x-ray-diffraction-analysis-of-a-mammal-inositol-1-3-4-5-6-pentakisphosphate-2-kinase
#17
Elsa Franco-Echevarría, Julia Sanz-Aparicio, Nathalie Troffer-Charlier, Arnaud Poterszman, Beatriz González
Inositol 1,3,4,5,6-pentakisphosphate 2-kinase (IP5 2-K) is an enzyme that catalyses the formation of phytic acid (IP6) from IP5 and ATP. In mammals, IP6 is involved in multiple events such as DNA repair and mRNA edit and it is the precursor of inositol pyrophosphates, emerging compounds shown to have an essential role in apoptosis. In addition, IP5 2-K have functions in cells independently of its catalytic activity, for example in rRNA biogenesis. We pursue the structure determination of a mammal IP5 2-K by Protein Crystallography...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28425008/reversible-activation-of-halophilic-%C3%AE-lactamase-from-methanol-induced-inactive-form-contrast-to-irreversible-inactivation-of-non-halophilic-counterpart
#18
Hiroko Tokunaga, Junpei Maeda, Tsutomu Arakawa, Masao Tokunaga
Effects of a water-miscible organic solvent, methanol, on the structure and activity of halophilic β-lactamase derived from Chromohalobacter sp.560 (HaBla), were investigated by means of circular dichroism (CD) measurement and enzymatic activity determination. Beta-lactamase activity was enhanced about 1.2-fold in the presence of 10-20% methanol. CD measurement of HaBla revealed different structures depending on the methanol concentration: native-like active form (Form I) in 10-20% methanol and methanol-induced inactive form at higher concentration (Form II in 40-60% and Form III in 75-80% methanol)...
June 2017: Protein Journal
https://www.readbyqxmd.com/read/28421415/a-tyrosine-residue-along-with-a-glutamic-acid-of-the-omega-like-loop-governs-the-beta-lactamase-activity-of-msmeg_4455-in-mycobacterium-smegmatis
#19
Ankita Bansal, Debasish Kar, Satya Deo Pandey, Ashok Matcha, N Ganesh Kumar, Soshina Nathan, Anindya S Ghosh
Mycobacterial beta-lactamases are involved in exerting beta-lactam resistance, though many of these proteins remain uncharacterized. Here, we have characterized MSMEG_4455 of Mycobacterium smegmatis as a beta-lactamase using molecular, biochemical and mutational techniques. To elucidate its nature in vivo and in vitro, and to predict its structure-function relationship in silico analysis is done. The MSMEG_4455 is cloned and expressed ectopically in a beta-lactamase deficient Escherichia coli mutant to establish the in vivo beta-lactamase like nature via minimum inhibitory concentration (MIC) determination...
June 2017: Protein Journal
https://www.readbyqxmd.com/read/28421414/polyclonal-antibodies-in-microplates-to-predict-the-maximum-adsorption-activities-of-enzyme-mutants-from-cell-lysates
#20
Yiran Feng, Xiaolan Yang, Deqiang Wang, Xiaolei Hu, Huimin Chong, Juan Liao, Chang-Guo Zhan, Fei Liao
With microplate-immobilized polyclonal antibodies against a starting enzyme or its active mutant bearing consistent accessible epitopes, the maximum activity of an adsorbed enzyme/mutant (Vs) was predicted for comparison to recognize weakly-positive mutants. Rabbit antisera against Escherichia coli alkaline phosphatase (ECAP) were fractionated with 33% ammonium sulfate to yield crude polyclonal antibodies for conventional immobilization in 96-well microplates. The response curve of the activities of ECAP/mutant adsorbed by the immobilized polyclonal antibodies to protein quantities from a cell lysate was fit to an approximation model to predict Vs...
June 2017: Protein Journal
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