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Resonance assignments of the human AKAP13-PH domain and stabilizing DH helix

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Abstract

The human AKAP13 protein contains DH and PH domains, which are responsible for its cell transforming activity. Despite its biomedical importance, the contribution of the PH domain to AKAP13 activity remains unclear and no three dimensional structure is available to date. Here we report the backbone and side chain 1H, 13C and 15N resonance assignments of a 20 kDa construct comprising the uniformly 13C and 15N labeled AKAP13-PH domain and an associated helix from the DH domain which is required for its stable expression. Resonance assignment has been achieved using conventional triple resonance experiments; 95% of all back bone resonances and more than 90% of side chain resonances have been successfully assigned. The 1H, 13C and 15N chemical shifts have been deposited in BMRB with accession number of 16195.

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Abbreviations

AKAP13:

A kinase anchor protein 13

Brx:

Breast cancer nuclear receptor-binding auxiliary protein

DH:

Dbl homology

EDTA:

Ethylenediaminetetraacetic acid

GEF:

Guanine nucleotide exchange factor

Lbc:

Lymphoid blast crisis

PH domain:

Pleckstrin homology

PKA:

Protein kinase A

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Acknowledgments

We thank all the staff of The Henry Wellcome Building for Biological NMR Spectroscopy, which is funded by the Wellcome Trust. This work was funded by Cancer Research UK and EU PRISM [M.O.] and carried out in collaboration with the Structural Genomics Consortium, a registered charity (number 1097737) funded by the Wellcome Trust, GlaxoSmithKline, Genome Canada, the Canadian Institutes of Health Research, the Ontario Innovation Trust, the Ontario Research and Development Challenge Fund and the Canadian Foundation for Innovation.

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Authors and Affiliations

  1. School of Cancer Sciences, University of Birmingham, Birmingham, B15 2TT, UK

    Masae Sugawara, Sara B.-M. Whittaker & Michael Overduin

  2. Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Str. 10, 13125, Berlin, Germany

    Linda Ball

  3. Structural Genomics Consortium, University of Oxford, Botnar Research Centre, Oxford, OX3 7LD, UK

    Shurene Bishop & Linda Ball

Authors
  1. Masae Sugawara
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  2. Sara B.-M. Whittaker
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  3. Shurene Bishop
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  4. Linda Ball
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  5. Michael Overduin
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Correspondence to Masae Sugawara.

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Sugawara, M., Whittaker, S.BM., Bishop, S. et al. Resonance assignments of the human AKAP13-PH domain and stabilizing DH helix. Biomol NMR Assign 3, 215–218 (2009). https://doi.org/10.1007/s12104-009-9178-0

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  • DOI: https://doi.org/10.1007/s12104-009-9178-0

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