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Crystal structure of wild-type centrin 1 from Mus musculus occupied by Ca2+

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Abstract

Mus musculus centrin 1 (MmCen1) is located at the cilium of photoreceptor cells connecting the outer segment through signal transduction components to the metabolically active inner segment. In the cilium, MmCen1 is involved in the translocation of transducin between compartments as a result of photoreceptor activation. In this study, we report the crystal structure of wild-type MmCen1 and its Ca2+-binding properties using structure-based mutagenesis. The crystal structure exhibits three structural features, i.e. four Ca2+ equally occupied at each EF-hand motif, structural changes accompanying helix motion at the N- and C-lobes, and adoption of N–C type dimerization when Ca2+ binds to EF-hand I and II in the N-lobe. The presence of MmCen1 dimers was confirmed in solution by native PAGE. Isothermal titration calorimetry data showed sequential binding of Ca2+ at four independent sites. Mutations S45A and D49A in EF-hand I alone disrupted the Ca2+-binding property of the wild-type protein. Based on the crystal structure of MmCen1, we suggest that a dimerization mode between the N- and C-lobes may be required by Ca2+ binding at the N-lobe.

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Abbreviations

CaM:

calmodulin

ITC:

isothermal titration calorimetry

MmCen1:

Mus musculus centrin 1

RIS:

rod inner segment

ROS:

rod outer segment

SPB:

spindle pole body

TnC:

troponin C

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Authors and Affiliations

  1. College of Environmental & Bioresource Sciences, Division of Biotechnology, Chonbuk National University, 54596, Iksan, Republic of Korea

    So Young Kim, Da Som Kim, Joo Eun Hong & Jung Hee Park

  2. Advanced Institute of Environment and Bioscience, Chonbuk National University, 54596, Iksan, Republic of Korea

    Jung Hee Park

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  1. So Young Kim
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  2. Da Som Kim
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Correspondence to Jung Hee Park.

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Published in Russian in Biokhimiya, 2017, Vol. 82, No. 10, pp. 1475-1488.

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Kim, S.Y., Kim, D.S., Hong, J.E. et al. Crystal structure of wild-type centrin 1 from Mus musculus occupied by Ca2+ . Biochemistry Moscow 82, 1129–1139 (2017). https://doi.org/10.1134/S0006297917100054

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  • DOI: https://doi.org/10.1134/S0006297917100054

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