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 journalArticle

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

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Atomic Force Microscopy
Hyaluronic Acid
Pasteurella multocida
Hyaluronic acid
Elongation
Atomic force microscopy
Polymers
Lipid bilayers
Enzymes
Mica
Lipid Bilayers
Cats
Uridine Diphosphate Glucuronic Acid
Uridine Diphosphate N-Acetylglucosamine
Uridine Diphosphate
Acetylglucosamine
Radioisotopes
Tail
Rate constants
Monomers

All Science Journal Classification (ASJC) codes

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

Cite this

Single-molecular enzymatic elongation of hyaluronan polymers visualized by high-speed atomic force microscopy. / Mori, Toshiaki; Hirose, Atsushi; Hagiwara, Tatsuya; Ohtsuka, Masanori; Kakuta, Yoshimitsu; Kimata, Koji; Okahata, Yoshio.

In: Journal of the American Chemical Society, Vol. 134, No. 50, 19.12.2012, p. 20254-20257.

Research output: Contribution to journalArticle

Mori, Toshiaki ; Hirose, Atsushi ; Hagiwara, Tatsuya ; Ohtsuka, Masanori ; Kakuta, Yoshimitsu ; Kimata, Koji ; Okahata, Yoshio. / Single-molecular enzymatic elongation of hyaluronan polymers visualized by high-speed atomic force microscopy. In: Journal of the American Chemical Society. 2012 ; Vol. 134, No. 50. pp. 20254-20257.
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AU - Kakuta, Yoshimitsu

AU - Kimata, Koji

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