A synthetic protocol for the fabrication of ultrathin polymeric films containing the enzyme 2-deoxy-d-ribose-5-phosphate aldolase from Escherichia coli (DERA) is presented. Ultrathin enzymatically active films are useful for applications in which only small quantities of active material are needed and at the same time quick response and contact times without diffusion limitation are wanted. We show how DERA as an exemplary enzyme can be immobilized in a thin polymer layer at the air-water interface and transferred to a suitable support by the Langmuir-Schaefer technique under full conservation of enzymatic activity. The polymer in use is a poly(N-isopropylacrylamide-co-N-2-thiolactone acrylamide) (P(NIPAAm-co-TlaAm)) statistical copolymer in which the thiolactone units serve a multitude of purposes including hydrophobization of the polymer, covalent binding of the enzyme and the support and finally cross-linking of the polymer matrix. The application of this type of polymer keeps the whole approach simple as additional cocomponents such as cross-linkers are avoided.
Pepper
is a fluorogenic RNA aptamer tag that binds to a variety
of benzylidene-cyanophenyl (HBC) derivatives with tight affinity and
activates their fluorescence. To investigate how Pepper RNA folds
to create a binding site for HBC, we used antibody-assisted crystallography
to determine the structures of Pepper bound to HBC530 and HBC599 to
2.3 and 2.7 Å resolutions, respectively. The structural data
show that Pepper folds into an elongated structure and organizes nucleotides
within an internal bulge to create the ligand binding site, assisted
by an out-of-plane platform created by tertiary interactions with
an adjacent bulge. As predicted from a lack of K+ dependence,
Pepper does not use a G-quadruplex to form a binding pocket for HBC.
Instead, Pepper uses a unique base-quadruple·base-triple stack
to sandwich the ligand with a U·G wobble pair. Site-bound Mg2+ ions support ligand binding structurally and energetically.
This research provides insight into the structural features that allow
the Pepper aptamer to bind HBC and show how Pepper’s function
may expand to allow the in vivo detection of other small molecules
and metals.
In the version of this Article originally published, the diblock copolymer structure in Fig. 2a showed a single bond between the carbon and the oxygen atoms; it should have been a double bond. This has been corrected in all versions of the Article.
Although many DNA nanodevices have been designed and synthesized to this day, a recent paper published in Nature Chemistry shows that the founder of structural DNA nanotechnology has developed the first 3D DNA crystal that can operate as a multi-state, reusable device. Ever since it was realized that DNA is the molecular code for life on Earth, it has been subjected to extensive and intensifying research and investigation. These studies range from understanding how four simple nucleotides encode information for every structure, directive, and procedure that keeps an organism viable to understanding the properties of the DNA polymer itself. The field of DNA nanotechnology encompasses a broad swath of these investigations, whereby DNA is being used to encode precise structures and responsive processes, recognition systems, selfassembly, and mechanical devices, among others. Here, we exhibit recent work by Seeman and colleagues in Nature Chemistry, where they have demonstrated the first synthetic DNA crystal that is a fully operational multistate device while in the solid crystalline state. 1
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.