Advances in amyloid research rely on improved access to the β-amyloid peptide, Aβ. N-Terminal methionine-extended Aβ, Aβ(M1-42), is a readily expressed and widely used form of Aβ with properties comparable to those of the natural Aβ(1-42) peptide. Expression of Aβ(M1-42) is simple to execute and avoids an expensive and often difficult enzymatic cleavage step associated with expression and isolation of Aβ(1-42). This paper reports an efficient method for the expression and purification of Aβ(M1-42) and N-labeled Aβ(M1-42). This method affords the pure peptide at ∼19 mg/L of bacterial culture through simple and inexpensive steps in 3 days. This paper also reports a simple method for the construction of recombinant plasmids and the expression and purification of Aβ(M1-42) peptides containing familial mutations. We anticipate that these methods will enable experiments that would otherwise be hindered by insufficient access to Aβ.
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http://dx.doi.org/10.1021/acs.biochem.8b00393 | DOI Listing |
STAR Protoc
January 2025
Department of Molecular Medicine, University of Pavia, Pavia, Italy. Electronic address:
Voltage-dependent anion channel 1 (VDAC1) is a key protein in cellular metabolism and apoptosis. Here, we present a protocol to express and purify milligram amounts of recombinant VDAC1 in Escherichia coli. We detail steps for a fluorescence polarization-based high-throughput screening assay using NADH displacement, along with procedures for thermostability, fluorescence polarization, and X-ray crystallography.
View Article and Find Full Text PDFPlanta
January 2025
Institute of Plant Genetics and Biotechnology, Plant Science and Biodiversity Center, Slovak Academy of Sciences, Akademicka 2, P. O. Box 39A, 950 07, Nitra, Slovak Republic.
DbChitI-3, Drosera binata's acidic chitinase, peaks at pH 2.5 from 15 °C to 30 °C. Gene expression is stimulated by polysaccharides and suppressed by monosaccharide digestion, implying a feedback loop in its transcriptional regulation.
View Article and Find Full Text PDFCell Mol Biol (Noisy-le-grand)
January 2025
Plant Protection Department, College of Food and Agriculture Sciences, King Saud University, P.O. Box 2460, Riyadh, 11451, Saudi Arabia.
Forty-four samples of garlic plants showing virus-like symptoms were collected, during the growing season (2021-2022) from different locations in Qassim province, Saudi Arabia. These samples were analyzed by ELISA against the important Allium allexiviruses including garlic virus A (GarV-A), garlic virus B (GarV-B), garlic virus C (GarV-C), and Shallot virus X (ShVX). The obtained results showed that 22/44 (50%) samples were found to be infected with one of the tested viruses.
View Article and Find Full Text PDFPak J Pharm Sci
January 2025
Department of Endocrinology, Gongli Hospital of Shanghai Pudong New Area, School of Gongli Hospital Medical Technology, University of Shanghai for Science and Technology, Shanghai, China.
Brown adipose tissue (BAT) is an ideal target organ for obesity treatment. Resinacein S is extracted from Ganoderma lucidum and can elevate Uncoupling protein 1 (UCP1) in cells, but its related effects at the animal level are not clear. The mice were fed with high-fat diet to construct obesity models and treated with Resinacein S.
View Article and Find Full Text PDFBiotechnol Bioeng
January 2025
Chair of Technical Biochemistry, Technische Universität Dresden, Dresden, Saxony, Germany.
Ikarugamycin is a member of the natural product family of the polycyclic tetramate macrolactams (PoTeMs). The compound exhibits a diverse range of biological activities, including antimicrobial, antiprotozoal, anti-leukemic, and anti-inflammatory properties. In addition, it interferes with several crucial cellular functions, such as oxidized low-density lipoprotein uptake in macrophages, Nef-induced CD4 cell surface downregulation, and mechanisms of endocytosis.
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