Binding properties of recombinant LDL receptor and LOX-1 receptor to LDL measured using bio-layer interferometry and atomic force microscopy.

Oxidation of low-density lipoproteins (LDLs) triggers a recognition by scavenger receptors such as lectin-like oxidized LDL receptor-1 (LOX-1) and is related to inflammation and cardiovascular diseases. Although LDLs that are recognized by LOX-1 can be risk-related LDLs, conventional LDL detection methods using commercially available recombinant receptors remain undeveloped. Using a bio-layer interferometry (BLI), we investigated the binding of recombinant LOX-1 (reLOX-1) and LDL receptors to the oxidized LDLs. The recombinant LDL receptor preferably bound minimally modified LDLs, while the reLOX-1 recognized extensively oxidized LDLs. An inversed response of the BLI was observed during the binding in the case of reLOX-1. AFM study showed that the extensively oxidized LDLs and aggregates of LDLs were observed on the surface, supporting the results. Altogether, a combined use of these recombinant receptors and the BLI method is useful in detecting high-risk LDLs such as oxidized LDLs and modified LDLs.

Histone chaperone-mediated co-expression assembly of tetrasomes and nucleosomes.

The nucleosome, a basic unit of chromatin found in all eukaryotes, is thought to be assembled through the orchestrated activity of several histone chaperones and chromatin assembly factors in a stepwise manner, proceeding from tetrasome assembly, to H2A/H2B deposition, and finally to formation of the mature nucleosome. In this study, we demonstrate chaperone-mediated assembly of both tetrasomes and nucleosomes on the well-defined Widom 601 positioning sequence using a co-expression/reconstitution wheat germ cell-free system. The purified tetrasomes and nucleosomes were positioned around the center of a given sequence. The heights and diameters were measured by atomic force microscopy. Together with the reported unmodified native histones produced by the wheat germ cell-free platform, our method is expected to be useful for downstream applications in the field of chromatin research.