caa a22 4500 397556187 CHVBK 20180308164802.0 cr unu---uuuuu 161202e199606 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021343 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021343 Innocuous Labeling of the Subfragment-2 Region of Skeletal Muscle Heavy Meromyosin with a Fluorescent Polyacrylamide Nanobead and Visualization of Individual Heavy Meromyosin Molecules [Elektronische Daten] [Yuki Kunioka, Toshio Ando] We have studied transglutaminase-catalyzed incorporation of monodansylcadaverine and monobiotincadaverine into rabbit skeletal muscle heavy meromyosin (HMM). The incorporation of dansylcadaverine reached saturation at 4 mol per 1 mol of HMM. An electrophoretogram of the chymotryptic digest of the dansyl-labeled HMM revealed that the labeling took place primarily in the S-2 region of HMM. Atomic force microscopic images and electron micrographs of the complexes of the biotinylated HMM and UltraAvidin-coated fluorescent polyacrylamide nanoparticles revealed that the biotinylated site on S-2 was very close to the C-terminus (near the S-2/light meromyosin junction). In keeping with this result, together with HMM's key sites being localized on the S-1 region, the enzymatic conjugation of biotincadaverine had no influence upon the actin-activated ATPase activity of HMM or upon the ability of HMM to actuate sliding of actin filaments in in vitro motility assay. Attachment of an UltraAvidin-coated fluorescent nanobead to the biotinylated HMM also did not alter the motile activity of HMM. Thus, we can optically pinpoint individual HMM molecules in a sample, which will facilitate handling and manipulation of single HMM molecules and observation of their functional behavior. © by the Journal of Biochemistry Regular Papers nationallicence actin nationallicence avidin nationallicence biotin nationallicence fluorescent nanobead nationallicence myosin nationallicence Kunioka Yuki Department of Physics, Faculty of Science, Kanazawa University, Kanazawa, Ishikawa 920-11 aut Ando Toshio Department of Physics, Faculty of Science, Kanazawa University, Kanazawa, Ishikawa 920-11 aut The Journal of Biochemistry Oxford University Press 119/6(1996-06), 1024-1032 0021-924X 119:6<1024 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021343 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021343 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kunioka Yuki Department of Physics, Faculty of Science, Kanazawa University, Kanazawa, Ishikawa 920-11 aut NATIONALLICENCE 700 1- Ando Toshio Department of Physics, Faculty of Science, Kanazawa University, Kanazawa, Ishikawa 920-11 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/6(1996-06), 1024-1032 0021-924X 119:6<1024 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford