Single-molecular enzymatic elongation of hyaluronan polymers visualized by high-speed atomic force microscopy

Toshiaki Mori, Atsushi Hirose, Tatsuya Hagiwara, Masanori Ohtsuka, Yoshimitsu Kakuta, Koji Kimata, Yoshio Okahata

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

Using high-speed scanning atomic force microscopy, we directly observed single-molecular enzymatic elongation of hyaluronan polymer chains at intervals of 10 s on a mica or lipid bilayer surface, on which Pasteurella multocida hyaluronic acid synthase (pmHAS) was immobilized. The reaction was started by the addition of both UDP-glucuronic acid and UDP-N-acetylglucosamine monomers. The average catalytic elongation rate constant (kcat) was found to be 1.8 mer s-1 from one active enzyme physically adsorbed on a mica surface. When pmHAS was immobilized by inserting its hydrophobic tail part into lipid bilayers, most of the enzymes retained their activity, and the k cat values were found to be in the range 1-10 mer s-1 for 29 enzymes (average was kcat = 2-4 mer s-1). These k cat values were lowest level of kcat = 1-100 s -1 obtained in bulk solution by radioisotope methods.

Original languageEnglish
Pages (from-to)20254-20257
Number of pages4
JournalJournal of the American Chemical Society
Volume134
Issue number50
DOIs
Publication statusPublished - Dec 19 2012

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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