Atomically flat mica surfaces were chemically modified with an alkyl trifluoromethyl ketone, a covalent inhibitor of esterase 2 from Alicyclobacillus acidocaldarius, which served as a tag for ligand-directed immobilization of esterase-linked proteins. Purified NADH oxidase from Thermus thermophilus and human exportin-t from cell lysates were anchored on the modified surfaces. The immobilization effectiveness of the proteins was studied by atomic force microscopy (AFM). It was shown that ligand-esterase interaction allowed specific attachment of exportin-t and resulted in high-resolution images and coverage patterns that were comparable with immobilized purified protein. Moreover, the biological functionality of immobilized human exportin-t in forming a quaternary complex with tRNA and the GTPase Ran-GTP, and the dimension changes before and after complex formation were also determined by AFM.
- MeSH
- bakteriální proteiny MeSH
- esterasy MeSH
- exprese genu MeSH
- financování organizované MeSH
- ketony farmakologie chemie MeSH
- lidé MeSH
- ligandy MeSH
- mikroskopie atomárních sil MeSH
- multienzymové komplexy genetika chemie MeSH
- NADH, NADPH oxidoreduktasy genetika chemie MeSH
- nukleocytoplazmatické transportní proteiny genetika chemie MeSH
- rekombinantní fúzní proteiny genetika chemie ultrastruktura MeSH
- silikáty hliníku chemie MeSH
- terciární struktura proteinů MeSH
- Thermus thermophilus enzymologie MeSH
- vazebná místa MeSH
- Check Tag
- lidé MeSH
