Bioorthogonal chemistry for selective recognition, separation and killing bacteria over mammalian cells

2016 ◽  
Vol 52 (17) ◽  
pp. 3482-3485 ◽  
Author(s):  
Zhenhua Li ◽  
Zhen Liu ◽  
Zhaowei Chen ◽  
Enguo Ju ◽  
Wei Li ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Mammalian Cells ◽  
Selective Recognition ◽  
Bioorthogonal Chemistry ◽  
New Strategy

We report a new strategy for selective recognition, separation and killing bacteria using metabolic engineering and bioorthogonal chemistry.

scholarly journals A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling

2021 ◽  
Vol 9 ◽  
Author(s):  
Birthe Meineke ◽  
Johannes Heimgärtner ◽  
Alexander J. Craig ◽  
Michael Landreh ◽  
Lindon W. K. Moodie ◽  
...  
Keyword(s):  
Amino Acids ◽  
Mammalian Cells ◽  
Stop Codon ◽  
Bioorthogonal Chemistry ◽  
Bioorthogonal Reaction ◽  
Noncanonical Amino Acids ◽  
Selective Reactivity ◽  
Direct Incorporation ◽  
Chelating Groups ◽  
Amber Suppression

Bioorthogonal chemistry allows rapid and highly selective reactivity in biological environments. The copper-catalyzed azide–alkyne cycloaddition (CuAAC) is a classic bioorthogonal reaction routinely used to modify azides or alkynes that have been introduced into biomolecules. Amber suppression is an efficient method for incorporating such chemical handles into proteins on the ribosome, in which noncanonical amino acids (ncAAs) are site specifically introduced into the polypeptide in response to an amber (UAG) stop codon. A variety of ncAA structures containing azides or alkynes have been proven useful for performing CuAAC chemistry on proteins. To improve CuAAC efficiency, biologically incorporated alkyne groups can be reacted with azide substrates that contain copper-chelating groups. However, the direct incorporation of copper-chelating azides into proteins has not been explored. To remedy this, we prepared the ncAA paz-lysine (PazK), which contains a picolyl azide motif. We show that PazK is efficiently incorporated into proteins by amber suppression in mammalian cells. Furthermore, PazK-labeled proteins show improved reactivity with alkyne reagents in CuAAC.

scholarly journals Design and application of circular RNAs with protein-sponge function

2020 ◽  
Vol 48 (21) ◽  
pp. 12326-12335
Author(s):  
Silke Schreiner ◽  
Anna Didio ◽  
Lee-Hsueh Hung ◽  
Albrecht Bindereif
Keyword(s):  
Alternative Splicing ◽  
Mammalian Cells ◽  
Regulatory Networks ◽  
Rna Binding ◽  
Molecular Medicine ◽  
Circular Rnas ◽  
Ca Repeat ◽  
New Strategy ◽  
Nuclear Ribonucleoprotein ◽  
Splicing Patterns

Abstract Circular RNAs (circRNAs) are a class of noncoding RNAs, generated from pre-mRNAs by circular splicing of exons and functionally largely uncharacterized. Here we report on the design, expression, and characterization of artificial circRNAs that act as protein sponges, specifically binding and functionally inactivating hnRNP (heterogeneous nuclear ribonucleoprotein) L. HnRNP L regulates alternative splicing, depending on short CA-rich RNA elements. We demonstrate that designer hnRNP L-sponge circRNAs with CA-repeat or CA-rich sequence clusters can efficiently and specifically modulate splicing-regulatory networks in mammalian cells, including alternative splicing patterns and the cellular distribution of a splicing factor. This new strategy can in principle be applied to any RNA-binding protein, opening up new therapeutic strategies in molecular medicine.

Multifunctional Cationic Poly(p-phenylene vinylene) Polyelectrolytes for Selective Recognition, Imaging, and Killing of Bacteria Over Mammalian Cells

2011 ◽  
Vol 23 (41) ◽  
pp. 4805-4810 ◽  
Author(s):  
Chunlei Zhu ◽  
Qiong Yang ◽  
Libing Liu ◽  
Fengting Lv ◽  
Shayu Li ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Selective Recognition ◽  
Phenylene Vinylene ◽  
Recognition Imaging

scholarly journals In Situ-Induced Multivalent Anticancer Drug Clusters in Cancer Cells for Enhancing Drug Efficacy

CCS Chemistry ◽  
2019 ◽  
pp. 97-105 ◽  
Author(s):  
Lingyun Zhou ◽  
Fengting Lv ◽  
Libing Liu ◽  
Shu Wang
Keyword(s):  
Cancer Cells ◽  
Anticancer Drug ◽  
Mammalian Cells ◽  
Hydrophobic Interactions ◽  
Drug Efficacy ◽  
Normal Cells ◽  
Cancer Chemotherapeutics ◽  
Drug Conjugates ◽  
Action Mechanisms ◽  
New Strategy

Increasing intracellular drug concentration is an effective way for cancer chemotherapeutics to enhance efficacy and combat drug resistance. In this work, a series of anticancer drug conjugates were prepared by linking thiol-modified oligo( p-phenylene vinylene) with paclitaxel, vincristine, teniposide, tamoxifen, doxorubicin, or podophyllotoxin (OPV-S-Drugs) through a Michael addition reaction. These OPV-S-Drugs could undergo intracellular assembly and aggregation upon oxidation to yield multivalent anticancer drug clusters, which inhibited their diffusion from cancer cells. The intracellular aggregation of OPV-S-Drugs originates from π–π stacking and hydrophobic interactions between OPV backbones, followed by cross-linking via disulfide bond formation in the presence of reactive oxygen species (ROS). The drug clusters occur only in the cytoplasm of cancer cells expressing high ROS levels, but not in healthy mammalian cells, thus reducing the cytotoxicity to normal cells. Specifically, the super-toxicity of podophyllotoxin to normal cells was obviously suppressed while the drug efficacy was maintained through our new strategy. The diverse action mechanisms of OPV-S-Drugs toward cancer cells is proposed.

scholarly journals A general strategy to construct small molecule biosensors in eukaryotes

eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
Justin Feng ◽  
Benjamin W Jester ◽  
Christine E Tinberg ◽  
Daniel J Mandell ◽  
Mauricio S Antunes ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Ligand Binding ◽  
Small Molecules ◽  
Small Molecule ◽  
Mammalian Cells ◽  
Fluorescent Protein ◽  
Dynamic Range ◽  
Plant Cells ◽  
General Strategy ◽  
Target Molecule

Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Here, we produce biosensors based on a ligand-binding domain (LBD) by using a method that, in principle, can be applied to any target molecule. The LBD is fused to either a fluorescent protein or a transcriptional activator and is destabilized by mutation such that the fusion accumulates only in cells containing the target ligand. We illustrate the power of this method by developing biosensors for digoxin and progesterone. Addition of ligand to yeast, mammalian, or plant cells expressing a biosensor activates transcription with a dynamic range of up to ~100-fold. We use the biosensors to improve the biotransformation of pregnenolone to progesterone in yeast and to regulate CRISPR activity in mammalian cells. This work provides a general methodology to develop biosensors for a broad range of molecules in eukaryotes.

scholarly journals Profiling of Glycosphingolipids with SCDase Digestion and HPLC-FLD-MS

2021 ◽  
Author(s):  
Radhika Chakraberty ◽  
Bela Reiz ◽  
Christopher Cairo
Keyword(s):  
T Cells ◽  
Fatty Acid ◽  
Anthranilic Acid ◽  
Mammalian Cells ◽  
Tissue Homogenate ◽  
Challenging Problem ◽  
Jurkat T Cells ◽  
External Standard ◽  
New Strategy ◽  
Cultured Human Cells

Lipid components of cells and tissues feature a large diversity of structures that present a challenging problem for molecular analysis. Glycolipids from mammalian cells contain glycosphingolipids (GSLs) as their major glycolipid component, and these structures vary in the identity of the glycan headgroup as well as the structure of the fatty acid and sphingosine (Sph) tails. Analysis of intact GSLs is challenging due to the low abundance of these species. Here, we develop a new strategy for the analysis of lyso-GSL (<i>l</i>-GSL), GSL that retain linkage of the glycan headgroup with the Sph base. The analysis begins with digestion of a GSL sample with sphingolipid ceramide <i>N</i>-deacylase (SCDase), followed by labelling with an amine-reactive fluorophore. The sample was then analyzed by HPLC-FLD-MS and quantitated by addition of an external standard. This method was compared analysis of GSL glycans after cleavage by an Endoglycoceramidase (EGCase) enzyme and labeling with a fluorophore (2-anthranilic acid, 2AA). The two methods are complementary, with EGCase providing improved signal (due to fewer species) and SCDase providing analysis of lyso-GSL. Importantly the SCDase method provides Sph composition of GSL species. We demonstrate the method on cultured human cells (Jurkat T cells) and tissue homogenate (porcine brain).

scholarly journals Synthesis and evaluation of Nα,Nε-diacetyl-l-lysine-inositol conjugates as cancer-selective probes for metabolic engineering of GPIs and GPI-anchored proteins

2020 ◽  
Vol 18 (15) ◽  
pp. 2938-2948 ◽  
Author(s):  
Mohit Jaiswal ◽  
Sanyong Zhu ◽  
Wenjie Jiang ◽  
Zhongwu Guo
Keyword(s):  
Metabolic Engineering ◽  
Cancer Cells ◽  
Cancer Targeting ◽  
Metabolic Labeling ◽  
Molecular Tools ◽  
New Strategy

This study has established a new strategy and new molecular tools for selective metabolic labeling of inositol on cancer cells, which should be useful for cancer targeting and study of GPI-anchored proteins.

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