Single-molecule analysis of chemoattractant-stimulated membrane recruitment of a PH-domain-containing protein

Journal of Cell Science
Satomi MatsuokaMasahiro Ueda

Abstract

Molecular mechanisms of chemotactic response are highly conserved among many eukaryotic cells including human leukocytes and Dictyostelium discoideum cells. The cells can sense the differences in chemoattractant concentration across the cell body and respond by extending pseudopods from the cell side facing to a higher concentration. Pseudopod formation is regulated by binding of pleckstrin homology (PH)-domain-containing proteins to phosphatidylinositol 3,4,5-trisphosphates [PtdIns(3,4,5)P3] localized at the leading edge of chemotaxing cells. However, molecular mechanisms underlying dynamic features of a pseudopod have not been fully explained by the known properties of PH-domain-containing proteins. To investigate the mechanisms, we visualized single molecules of green fluorescent protein tagged to Crac (Crac-GFP), a PH-domain-containing protein in D. discoideum cells. Whereas populations of Crac molecules exhibited a stable steady-state localization at pseudopods, individual molecules bound transiently to PtdIns(3,4,5)P3 for approximately 120 milliseconds, indicating dynamic properties of the PH-domain-containing protein. Receptor stimulation did not alter the binding stability but regulated the number of bound PH-domain mol...Continue Reading

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Citations

Apr 24, 2008·Annual Review of Biomedical Engineering·Jay T GrovesMartin B Forstner
Jan 18, 2013·PLoS Computational Biology·Satomi MatsuokaMasahiro Ueda
May 19, 2009·Experimental Animals·Hideko Urushihara
Sep 15, 2009·Seminars in Cell & Developmental Biology·Manli ChuaiCornelis J Weijer
Feb 19, 2008·Biochemical and Biophysical Research Communications·Guo FuZhi-Zhan Xu
Dec 23, 2006·Bio Systems·Masayuki J SatoMasahiro Ueda
Sep 14, 2010·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Pablo A Iglesias
Aug 11, 2006·The EMBO Journal·Dirk Dormann, Cornelis J Weijer
Mar 8, 2011·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Tian Jin
Jun 30, 2006·Current Opinion in Genetics & Development·Pascale G Charest, Richard A Firtel
Aug 19, 2009·Biophysical Journal·Satomi MatsuokaMasahiro Ueda
Jan 27, 2009·Biophysical Journal·Jefferson D Knight, Joseph J Falke
Sep 12, 2014·PLoS Computational Biology·Masato YasuiMasahiro Ueda
Aug 25, 2009·The Neuroscientist : a Review Journal Bringing Neurobiology, Neurology and Psychiatry·Kyonsoo Hong, Makoto Nishiyama
Mar 26, 2013·Integrative Biology : Quantitative Biosciences From Nano to Macro·Miguel CoelhoIva M Tolić-Nørrelykke
Feb 1, 2019·Communications Biology·Hidenori HashimuraMasahiro Ueda
Oct 28, 2018·Nature Communications·Satomi Matsuoka, Masahiro Ueda
Feb 25, 2020·Microscopy·Michio HiroshimaMasahiro Ueda
Sep 14, 2011·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Yasushi SakoAkihiro Yamamoto
Jan 12, 2020·Communications Biology·Hidenori HashimuraMasahiro Ueda
Mar 3, 2012·Cold Spring Harbor Protocols·Satomi MatsuokaMasahiro Ueda

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