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J. Biol. Chem., Vol. 283, Issue 4, 1848-1856, January 25, 2008

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Two Stage Cadherin Kinetics Require Multiple Extracellular Domains but Not the Cytoplasmic Region^*

Yuan-Hung Chien, Ning Jiang¹, Fang Li^¶, Fang Zhang, Cheng Zhu², and Deborah Leckband^||3

From the Department of Biochemistry, University of Illinois, Urbana, Illinois, the Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, ^¶Theoretical and Applied Mechanics, University of Illinois, Urbana, Illinois, and the ^||Department of Chemistry, University of Illinois, Urbana, Illinois 61801

Micropipette manipulation measurements quantified the pre-steady state binding kinetics between cell pairs mediated by Xenopus cleavage stage cadherin. The time-dependence of the intercellular binding probability exhibits a fast forming, low probability binding state, which transitions to a slower forming, high probability state. The biphasic kinetics are independent of the cytoplasmic region, but the transition to the high probability state requires the third extracellular domain EC3. Deleting either EC3 or EC3–5, or substituting Trp² for Ala reduces the binding curves to a simple, monophasic rise in binding probability to a limiting plateau, as predicted for a single site binding mechanism. The two stage cadherin binding process reported here directly parallels previous biophysical studies, and confirms that the cadherin ectodomain governs the initial intercellular adhesion dynamics.

Received for publication, September 26, 2007 , and in revised form, November 5, 2007.

^* This work was supported in part by National Institutes of Health Grants R01 GM51338 (to D. E. L.) and by R01 AI38282 and R01 AI44902 (to C. Z.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Current address: Dept. of Bioengineering, Stanford University, Stanford, CA.

² To whom correspondence may be addressed: 315 Ferst Drive, Atlanta, GA 30332. Tel.: 404-894-3269; Fax: 404-385-1397; E-mail: cheng.zhu{at}bme.gatech.edu. ³ To whom correspondence may be addressed: 600 South Mathews Ave. Urbana, IL 61820. Tel.: 217-244-0793; Fax: 217-333-5052; E-mail: Leckband{at}uiuc.edu.

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