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A Metal-Chelating Lipid for 2D Protein Crystallization via Coordination of Surface Histidines

Pack, Daniel W. and Chen, Guohua and Maloney, Kevin M. and Chen, Chao-Tsen and Arnold, Frances H. (1997) A Metal-Chelating Lipid for 2D Protein Crystallization via Coordination of Surface Histidines. Journal of the American Chemical Society, 119 (10). pp. 2479-2487. ISSN 0002-7863. doi:10.1021/ja964099e.

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Two-dimensional protein crystallization on lipid monolayers is becoming a powerful technique for structure determination as well as materials applications. However, progress has been hindered by the requirement of a unique affinity lipid for each new protein of interest. Metal ion coordination by surface-accessible histidine side chains provides a convenient and general method for targeting of proteins to surfaces. Here we present the synthesis and characterization of a metal-chelating lipid which has been designed to target proteins to Langmuir monolayers and promote their two-dimensional crystallization based on histidine coordination. The lipid utilizes the metal chelator iminodiacetate (IDA) as the hydrophilic headgroup and contains unsaturated, oleyl tails to provide the fluidity necessary for two-dimensional protein crystallization. The lipid is shown to bind copper from the subphase strongly when incorporated in Langmuir monolayers. In addition, it is possible to form copper-containing monolayers by spreading the premetalated lipid on the subphase in the absence of copper. Fluorescence microscopy reveals the binding and crystallization of the protein streptavidin, promoted by the simultaneous coordination of two surface-accessible histidine side chains to the IDA−Cu lipid.

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Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 1997 American Chemical Society. Received November 27, 1996. Publication Date (Web): March 12, 1997. We thank Dr. Viola Vogel, Dr. Wolfgang Frey, and William R. Schief, Jr., University of Washington, Seattle, for helpful discussions. In addition, we thank Wei-Hau Chang and Dr. Roger D. Kornberg, at Stanford University, for performing the electron diffraction analysis of the 2D streptavidin crystals. This work was supported by the Army Research Office. D.W.P. acknowledges a Landau fellowship and a training fellowship from the National Institute of General Medical Sciences, NRSA Award 1 T32 GM 08346-01. G.C. and K.M.M. are supported by postdoctoral fellowships from the National Institute of General Medical Sciences, NRSA Awards 5 F32 GM17448-02 and 5 F32 GM16953-02, respectively.
Funding AgencyGrant Number
Army Research Office (ARO)UNSPECIFIED
NIH Predoctoral Fellowship1 T32 GM 08346-01
NIH Postdoctoral Fellowship5 F32 GM17448-02
NIH Postdoctoral Fellowship5 F32 GM16953-02
Issue or Number:10
Record Number:CaltechAUTHORS:20180801-090115992
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Official Citation:A Metal-Chelating Lipid for 2D Protein Crystallization via Coordination of Surface Histidines. Daniel W. Pack, Guohua Chen, Kevin M. Maloney, Chao-Tsen Chen, and, and Frances H. Arnold. Journal of the American Chemical Society 1997 119 (10), 2479-2487. DOI: 10.1021/ja964099e
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:88426
Deposited By: Tony Diaz
Deposited On:01 Aug 2018 16:14
Last Modified:16 Nov 2021 00:26

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