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Science 26 October 2001:
Vol. 294. no. 5543, pp. 864 - 867
DOI: 10.1126/science.1063951
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Reports

Single-Molecule Analysis of Chemotactic Signaling in Dictyostelium Cells

Masahiro Ueda,16* Yasushi Sako,26 Toshiki Tanaka,4dagger Peter Devreotes,5 Toshio Yanagida36

Single-molecule imaging techniques were used to reveal the binding of individual cyclic adenosine 3',5'-monophosphate molecules to heterotrimeric guanine nucleotide-binding protein coupled receptors on the surface of living Dictyostelium discoideum cells. The binding sites were uniformly distributed and diffused rapidly in the plane of the membrane. The probabilities of individual association and dissociation events were greater for receptors at the anterior end of the cell. Agonist-induced receptor phosphorylation had little effect on any of the monitored properties, whereas G protein coupling influenced the binding kinetics. These observations illustrate the dynamic properties of receptors involved in gradient sensing and suggest that these may be polarized in chemotactic cells.

1 Recognition and Formation,
2 Time's Arrow and Biosignaling, Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Corporation (JST).
3 Single Molecule Processes Project, International Cooperative Research Project (ICORP), JST, Mino, Osaka 562-0035, Japan.
4 Biomolecular Engineering Research Institute, Suita, Osaka 565-0874, Japan.
5 Department of Cell Biology, Johns Hopkins Medical Institutions, Baltimore, MD 21205, USA.
6 Department of Physiology and Biosignaling, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, Japan.
*   To whom correspondence should be addressed. E-mail: ueda{at}phys1.med.osaka-u.ac.jp

dagger    Present address: Department of Applied Chemistry, Nagoya Institute of Technology, Showa-ku, Nagoya 466-8555, Japan.

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