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Preliminary crystallization studies of calmodulin-dependent protein phosphatase (calcineurin) from bovine brain

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Abstract

Calcineurin is a serine/threonine protein phosphatase which catalyzes the hydrolysis of both phosphoseryl/phosphothreonyl and phosphotyrosyl proteins as well as low molecular weight compounds such as p-nitrophenyl phosphate. It is a hetero-dimeric protein consisting of a 60 kDa A chain and 19 kDa B chain. Calcineurin A is organized into functionally distinct domains such as a catalytic domain, a calcineurin B binding domain, a calmodulin-binding domain, and an inhibitory domain. Calcineurin B has four EF-hand calcium binding domains with a secondary structure that is homologous to calmodulin but its metal binding properties are more similar to troponin-C. The N-terminal myristoyl group of calcineurin B might play a role in the interaction between subunits A and B during phosphorylation/dephosphorylation processes. Crystals of size 0.125×0.07×0.03 mm and 0.7×0.03×0.02 mm have been obtained for calcineurin and the A subunit respectively. Crystals of calcineurin show strong diffraction to 5.3 Å and weak diffraction to 3.0 Å on rotating anode operated at 50 kV and 100 mA. Further work is in progress to improve the X-ray diffraction quality of these crystals and to obtain well diffracting crystals of calcineurin B.

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Authors and Affiliations

  1. Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, 3307 North Broad Street, Allied Health Building, Room 557, 19140, Philadelphia, PA, USA

    K. Balendiran & Krishna H. M. Murthy

  2. Department of Pathology and Saskatoon Cancer Center, Royal University Hospital, University of Saskatchewan, S7N OXO, Saskatoon, Saskatchewan, Canada

    Yingchun Tan & Rajendra K. Sharma

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  1. K. Balendiran
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  2. Yingchun Tan
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  3. Rajendra K. Sharma
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  4. Krishna H. M. Murthy
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Balendiran, K., Tan, Y., Sharma, R.K. et al. Preliminary crystallization studies of calmodulin-dependent protein phosphatase (calcineurin) from bovine brain. Mol Cell Biochem 149, 127–130 (1995). https://doi.org/10.1007/BF01076570

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