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Apurinic/apyrimidinic endonuclease 1 (APE1) is a multifunctional DNA repair protein localized in different subcellular compartments. The mechanisms responsible for the highly regulated subcellular localization and “interactomes” of this protein are not fully understood but have been closely correlated to the posttranslational modifications in different biological context. In this work, we attempted to develop a bio-nanocomposite with antibody-like properties that could capture APE1 from cellular matrices to enable the comprehensive study of this protein. By fixing the template APE1 on the avidin-modified surface of silica-coated magnetic nanoparticles, we first added 3-aminophenylboronic acid to react with the glycosyl residues of avidin, followed by addition of 2-acrylamido-2-methylpropane sulfonic acid as the second functional monomer to perform the first step imprinting reaction. To further enhance the affinity and selectivity of the binding sites, we carried out the second step imprinting reaction with dopamine as the functional monomer. After the polymerization, we modified the nonimprinted sites with methoxypoly (ethylene glycol) amine (mPEG-NH2). The resulting molecularly imprinted polymer-based bio-nanocomposite showed high affinity, specificity, and capacity for template APE1. It allowed for the extraction of APE1 from the cell lysates with high recovery and purity. Moreover, the bound protein could be effectively released from the bio-nanocomposite with high activity. The bio-nanocomposite offers a very useful tool for the separation of APE1 from various complex biological samples. 相似文献
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《Trends in biotechnology》2020,38(4):368-387
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Major intrinsic proteins (MIPs) in plants: a complex gene family with major impacts on plant phenotype 总被引:2,自引:0,他引:2
The ubiquitous cell membrane proteins called aquaporins are now firmly established as channel proteins that control the specific
transport of water molecules across cell membranes in all living organisms. The aquaporins are thus likely to be of fundamental
significance to all facets of plant growth and development affected by plant–water relations. A majority of plant aquaporins
have been found to share essential structural features with the human aquaporin and exhibit water-transporting ability in
various functional assays, and some have been shown experimentally to be of critical importance to plant survival. Furthermore,
substantial evidence is now available from a number of plant species that shows differential gene expression of aquaporins
in response to abiotic stresses such as salinity, drought, or cold and clearly establishes the aquaporins as major players
in the response of plants to conditions that affect water availability. This review summarizes the function and regulation
of these genes to develop a greater understanding of the response of plants to water insufficiency, and particularly, to identify
tolerant genotypes of major crop species including wheat and rice and plants that are important in agroforestry.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
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《Journal of molecular recognition : JMR》2017,30(1)
In recent years, there has been a tremendous increase in the papers published on synthetic recognition elements. Molecularly imprinted polymers (MIPs), also referred to as “man‐made mimics” of antibodies, are able to rebind their template molecules with high affinity. Advantages compared with those of natural receptors include their excellent thermal and chemical stability, low cost, and ease of the production process. However, their use in commercial biosensors is limited owing to the difficulty to incorporate MIPs into suitable sensing platforms and traditional detection techniques, such as chromatography, that require bulky and sophisticated equipment. In this review, we evaluate the potential to use MIPs combined with thermal read‐out for the detection of low‐weight organic molecules. We discuss thermal methods to study MIP‐template complexation and to determine neurotransmitters concentrations. In particular, we highlight the heat‐transfer method, a recent technique that is straightforward and low cost and requires minimal instrumentation. Until now, sample preparation involves a 2‐step process, making it time‐consuming, and measuring biological samples is difficult owing to the noise in the signal. Different sample preparation methods are discussed, and it will be demonstrated how this affects the thermal response. An outlook is given in novel methods that can simplify and speed up sample preparation. Finally, we show a novel thermal technique, which is based on the analysis of transport of thermal waves rather than evaluating the fixed heat‐transfer resistance. Through applying the concept of thermal waves, signal‐noise ratio is significantly increased, which results in lower detection limits and has potential for the study of biological samples. 相似文献
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Molecularly imprinted polymers (MIPs) are artificial antibodies for a target molecule. The review focuses mainly on mechanistic steps involved in forming MIPs and the role of co-monomers and porogen. In addition, the electronic transition between different energy levels is explained with the help of the Jablonski diagram. Diverse receptor and target molecules for anchoring artificial MIPs are discussed, accentuating the synergetic effects obtained. The binding efficiency, selectivity, and sensitivity of various optical sensors are discussed intensively. In addition to this, we focused on synthesis, physical forms, characterization techniques, and microorganism detection of imprinted polymers. A brief investigation on the use of MIPs in cancer diagnosis is also included, and attention is extended to the important challenges faced in using imprinted polymers. 相似文献
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