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Optimization of expression, purification, and NMR measurement for structural studies of transmembrane region of amyloid β protein

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Abstract

The amyloid precursor protein (APP) is an integral transmembrane protein which has been suggested to play a central role in the pathogenesis of Alzheimer’s disease. Despite the enormous amount of research conducted on amyloid protein, the precise mechanism of its toxic effect is not yet fully understood. To better understand the mechanism and function of amyloid protein, it is critical to elucidate the three-dimensional structure of the single transmembrane spanning region of human APP (hAPP-TM). Unfortunately, it is difficult to prepare the peptide sample because hAPP-TM is a membrane-bound protein that transverses the lipid bilayer of the cell membrane. Generally, the preparation of a transmembrane peptide is very difficult and time-consuming. In fact, high yield production of transmembrane peptides has been limited by experimental difficulties related to insufficient yields and the low solubility of such peptides. In this study, we describe experimental processes developed to optimize the expression, purification, and NMR measurement conditions for hAPP-TM transmembrane peptide.

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Authors and Affiliations

  1. Department of Chemistry and Protein Research Center for Bio-Industry, Hankuk University of Foreign Studies, Yongin, 449-791, Korea

    Tae-Joon Park, Ji-Sun Kim, Sung-Sub Choi & Yongae Kim

Authors
  1. Tae-Joon Park
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  2. Ji-Sun Kim
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  3. Sung-Sub Choi
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  4. Yongae Kim
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Correspondence to Yongae Kim.

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Park, TJ., Kim, JS., Choi, SS. et al. Optimization of expression, purification, and NMR measurement for structural studies of transmembrane region of amyloid β protein. Biotechnol Bioproc E 16, 477–481 (2011). https://doi.org/10.1007/s12257-010-0276-8

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  • DOI: https://doi.org/10.1007/s12257-010-0276-8

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