作者: Nonthreatening 時間: 2025-3-21 21:24
Engineering Homogeneous Photoactive Antibody Fragmentse describe a procedure for engineering photoactive antibody fragments that bind to their target antigen only after irradiation with 365?nm light. The procedure starts with identification of tyrosine residues in antibody fragments that are important for antibody–antigen binding and thus targets for r作者: jeopardize 時間: 2025-3-22 03:10
Repurposing Photosensitizer Proteins Through Genetic Code Expansion to Facilitate Photo-Biocatalysisalysts. Genetically encoded photo-redox cofactors endow photoenzymes with enhanced or novel activities that catalyze numerous transformations with high efficiency. Herein, we describe a protocol of repurposing photosensitizer proteins (.) through genetic code expansion to facilitate multiple photoca作者: SPASM 時間: 2025-3-22 08:07
Genetic Encoding of a Fluorescent Noncanonical Amino Acid as a FRET Donor for the?Analysis of Deubiqing of proteins. These co-translational and internal fluorescent tags have been harnessed to establish genetically encoded F?rster resonance energy transfer (FRET) probes for studying protein structural changes and interactions. Here, we describe the protocols for site-specific incorporation of an a作者: inchoate 時間: 2025-3-22 12:19
Creating Selenocysteine-Specific Reporters Using Inteins. These characteristics are attractive for designing highly active enzymes or extremely stable proteins and studying protein folding or electron transfer, to name a few. There are also 25 human selenoproteins, of which many are essential for our survival. The ability to create or study these selenop作者: 發(fā)酵劑 時間: 2025-3-22 13:36 作者: 發(fā)酵劑 時間: 2025-3-22 17:28 作者: 高談闊論 時間: 2025-3-23 00:40 作者: CLASH 時間: 2025-3-23 04:08
Genetically Encoded Noncanonical Amino Acids in Proteins to Investigate Lysine Benzoylationion and degradation. Histone lysine benzoylation is a recently identified epigenetic marker associated with active transcription, which has physiological relevance distinct from histone acetylation and can be regulated by debenzoylation of sirtuin 2 (SIRT2). Herein, we provide a protocol for the inc作者: 同步信息 時間: 2025-3-23 08:29
Semisynthesis of Glutamine-Methylated Proteins Enabled by Genetic Code Expansionelucidate the biological implications of this modification. Herein, we describe a protocol to generate histones with site-specific Gln methylation in a semisynthetic manner. Firstly, an esterified glutamic acid analogue (BnE) is genetically encoded into proteins by genetic code expansion with high e作者: Enervate 時間: 2025-3-23 12:27
Genetic Code Expansion in Mammalian Cellsch a unique handle into the protein of interest (POI), bioorthogonal reactions can be utilized in live cells to monitor or manipulate the interaction, translocation, function, and modification of the POI. Here, we describe a basic protocol outlining the necessary steps to incorporate a ncAA into a P作者: indicate 時間: 2025-3-23 14:49
Generation of Amber Suppression Cell Lines Using CRISPR-Cas9ing cell. The archaeal pyrrolysine-tRNA/pyrrolysine-tRNA synthetase (PylT/RS) pair from . (.) has been established for incorporation of a wide range of noncanonical amino acids (ncAAs) in mammalian cells. Once integrated in an engineered protein, ncAAs allow for simple click-chemistry derivatization作者: 無畏 時間: 2025-3-23 20:03 作者: white-matter 時間: 2025-3-23 23:12
Genetically Encoded 1,2-Aminothiol for Site-Specific Modification of a Cellular Membrane Protein viaion on a target protein is through a reaction between bioorthogonal functionalities. Indeed, various bioorthogonal reactions have been developed, including a recently reported reaction between 1,2-aminothiol and ((alkylthio)(aryl)methylene)malononitrile (TAMM). Here, we describe the procedure that c作者: Intuitive 時間: 2025-3-24 06:06 作者: 眉毛 時間: 2025-3-24 07:07 作者: 食道 時間: 2025-3-24 12:38
Site-Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cellsl processes and the development of human diseases, including AIDS and cancer. To facilitate the study of PTS in live mammalian cells, an approach for the site-specific synthesis of tyrosine-sulfated proteins (sulfoproteins) was developed. This approach takes advantage of an evolved . tyrosyl-tRNA sy作者: overshadow 時間: 2025-3-24 17:39
Book 2023ion provides a broad resource of methods for implementing GCE in .E. coli., mammalian cells, and animals, highlighting specific applications ranging from fluorescence labeling to photocontrol and the study of protein post-translational modification. Written for the highly successful .Methods in Mole作者: 名字 時間: 2025-3-24 22:01 作者: 嬰兒 時間: 2025-3-25 00:17 作者: ALE 時間: 2025-3-25 06:37 作者: 招募 時間: 2025-3-25 11:23 作者: 可卡 時間: 2025-3-25 14:41 作者: 發(fā)現(xiàn) 時間: 2025-3-25 18:02 作者: 浸軟 時間: 2025-3-25 20:59
Encoding Noncanonical Amino Acids into Phage Displayed Proteinsalities and building blocks in proteins displayed on phage provides the foundation for further phage display applications in fields such as imaging, protein targeting, and the production of new materials.作者: 嬉耍 時間: 2025-3-26 01:40
Conformational GPCR BRET Sensors Based on Bioorthogonal Labeling of Noncanonical Amino Acidsonal sensors based on the minimally invasive bioorthogonal labeling procedure, we describe a method that can be used in a microtiter plate format and can be easily adopted to investigate ligand-induced dynamics in various membrane receptors.作者: 伸展 時間: 2025-3-26 04:58 作者: 我不重要 時間: 2025-3-26 10:23
Fault-Tolerant System Technology,eplacement with photocaged tyrosine (pcY). This is followed by cloning of plasmids and expression of pcY-containing antibody fragments in .. Finally, we describe a cost-effective and biologically-relevant method for measuring the binding affinity of photoactive antibody fragments to antigens expressed on the surface of live cancer cells.作者: Sarcoma 時間: 2025-3-26 12:48
Nonrenewable Two-State Systems,minocoumarin-derived fluorescent ncAA into proteins in . and preparation of a fluorescent ncAA-based FRET probe for assaying the?activities of deubiquitinases, a key class of enzymes involved in ubiquitination. We also describe the deployment of an in vitro fluorescence assay to screen and analyze small-molecule inhibitors against deubiquitinases.作者: omnibus 時間: 2025-3-26 20:09 作者: Abduct 時間: 2025-3-27 00:02 作者: 抱負(fù) 時間: 2025-3-27 01:25
https://doi.org/10.1007/978-1-349-16014-3fficiency, which can be quantitatively converted into an acyl hydrazide via hydrazinolysis. Then, through a reaction with acetyl acetone, the acyl hydrazide is converted to reactive Knorr pyrazole. Finally, the in situ generated Knorr pyrazole is incubated with methylamine to give Gln methylation.作者: 投射 時間: 2025-3-27 08:08 作者: VICT 時間: 2025-3-27 10:44
Focused Engineering of Pyrrolysyl-tRNA Synthetase-Based Orthogonal Translation Systems for the Incors biotechnological and even therapeutically relevant applications. Here we present a protocol of engineering PylRS for novel substrates with unique chemical functionalities. These functional groups can act as intrinsic probes, especially in complex biological environments such as mammalian cells, tissues, and even whole animals.作者: Onerous 時間: 2025-3-27 14:19
Engineering Homogeneous Photoactive Antibody Fragmentseplacement with photocaged tyrosine (pcY). This is followed by cloning of plasmids and expression of pcY-containing antibody fragments in .. Finally, we describe a cost-effective and biologically-relevant method for measuring the binding affinity of photoactive antibody fragments to antigens expressed on the surface of live cancer cells.作者: Innocence 時間: 2025-3-27 19:32
Genetic Encoding of a Fluorescent Noncanonical Amino Acid as a FRET Donor for the?Analysis of Deubiqminocoumarin-derived fluorescent ncAA into proteins in . and preparation of a fluorescent ncAA-based FRET probe for assaying the?activities of deubiquitinases, a key class of enzymes involved in ubiquitination. We also describe the deployment of an in vitro fluorescence assay to screen and analyze small-molecule inhibitors against deubiquitinases.作者: JAUNT 時間: 2025-3-27 23:10 作者: 誰在削木頭 時間: 2025-3-28 05:28 作者: 不確定 時間: 2025-3-28 06:38 作者: 就職 時間: 2025-3-28 14:05 作者: cauda-equina 時間: 2025-3-28 16:06
1064-3745 expertsThis detailed volume explores non-canonical amino acids (ncAAs) through their site-specific incorporation by genetic code expansion (GCE). The collection provides a broad resource of methods for implementing GCE in .E. coli., mammalian cells, and animals, highlighting specific applications r作者: AIL 時間: 2025-3-28 22:31 作者: florid 時間: 2025-3-29 01:12 作者: 分貝 時間: 2025-3-29 05:26
Monitoring Your Containerized Environment,account of the incorporation of sTyr in HEK293T cells using the enhanced green fluorescent protein as an example. This method can be widely applied to incorporating sTyr into any POI to investigate the biological functions of PTS in mammalian cells.作者: 進(jìn)取心 時間: 2025-3-29 09:07
Repurposing Photosensitizer Proteins Through Genetic Code Expansion to Facilitate Photo-Biocatalysispurification, and characterization of the . are detailed. The installation of the catalytic modules and the utilization of .-based artificial photoenzymes for photoenzymatic CO. reduction and dehalogenation are also described.作者: Presbyopia 時間: 2025-3-29 14:03 作者: 使隔離 時間: 2025-3-29 16:54 作者: 頌揚(yáng)本人 時間: 2025-3-29 22:31 作者: 不能平靜 時間: 2025-3-30 02:55 作者: 光明正大 時間: 2025-3-30 08:04
Book 2023d Non-Canonical Amino Acids: Methods and Protocols. serves as an ideal source of methodologies that can be adapted and extended, migrated to different model systems, and combined in new ways to help explore a wide range of biological questions and to augment industrial and pharmaceutical protein engineering..作者: 賄賂 時間: 2025-3-30 08:45 作者: 邊緣 時間: 2025-3-30 12:24
https://doi.org/10.1007/978-1-4842-9277-8ion. Here, we demonstrate this method by using cAMP-dependent protein kinase catalytic subunit alpha (PKA-Cα) as the model enzyme. The method should be applicable to other kinases, enabling their rapid and selective inhibition.作者: Conserve 時間: 2025-3-30 18:16
Generation of Amber Suppression Cell Lines Using CRISPR-Cas9eaks (DSBs) and nonhomologous end joining (NHEJ) repair to target the PylT/RS expression cassette to the . safe harbor locus in human cells. .PylRS expression from this single locus is sufficient for efficient amber suppression when the cells are subsequently transfected transiently with a PylT/gene作者: Oscillate 時間: 2025-3-30 22:21 作者: debble 時間: 2025-3-31 04:26 作者: Flounder 時間: 2025-3-31 07:52
Fault-Tolerant System Technology,e describe a procedure for engineering photoactive antibody fragments that bind to their target antigen only after irradiation with 365?nm light. The procedure starts with identification of tyrosine residues in antibody fragments that are important for antibody–antigen binding and thus targets for r作者: 頭腦冷靜 時間: 2025-3-31 11:30
Dependability in Medicine and Neurologyalysts. Genetically encoded photo-redox cofactors endow photoenzymes with enhanced or novel activities that catalyze numerous transformations with high efficiency. Herein, we describe a protocol of repurposing photosensitizer proteins (.) through genetic code expansion to facilitate multiple photoca作者: 漂白 時間: 2025-3-31 14:01 作者: 有節(jié)制 時間: 2025-3-31 21:21 作者: Benign 時間: 2025-3-31 22:06
https://doi.org/10.1007/b102376oncanonical amino acids (ncAAs) by orthogonal aminoacyl-tRNA synthetase (aaRS)/tRNA pairs using nonsense codons, which could significantly expand new functionalities of proteins in both scientific and biomedical applications. Here, by hijacking the cysteine biosynthetic enzymes, we describe a method
