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Variability of calcium binding to EF-hand motifs probed by electrospray ionization mass spectrometry

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Journal of the American Society for Mass Spectrometry

Abstract

The modulation of calcium binding by the EF-hand motifs present in a calmodulin (CAM) homologue, a calcium binding protein (CaBP) from Entamoeba histolytica by three external parameters-pH, ligand coordinator EGTA, and fragmentor voltage was investigated by mass spectrometry. Calcium binding follows expected patterns at highly acidic and alkaline pH with the preponderance of the apo and the completely saturated forms, respectively. Surprisingly, additional nonspecific binding is observed near neutral pH. Studies on EGTA chelation and effects of fragmentor voltage showed cooperativity in calcium removal in at least one of the domains. Similar studies on a smaller construct containing the two high affinity carboxy terminal sites revealed interesting differences and provided an estimate of the specificity and tolerance of the EF-hand motifs to calcium binding and removal.

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Author notes

  1. B. Gopal

    Present address: Division of Mycobacterial Research and Protein Structure, National Institute for Medical Research, NW7 1AA, Mill Hill, London, UK

Authors and Affiliations

  1. Molecular Biophysics Unit, Indian Institute of Science, 560 012, Bangalore, India

    Anu K. Moorthy, S. K. Singh, B. Gopal, A. Surolia & M. R. N. Murthy

Authors
  1. Anu K. Moorthy
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  2. S. K. Singh
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  3. B. Gopal
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  4. A. Surolia
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  5. M. R. N. Murthy
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Correspondence to M. R. N. Murthy.

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Moorthy, A.K., Singh, S.K., Gopal, B. et al. Variability of calcium binding to EF-hand motifs probed by electrospray ionization mass spectrometry. J Am Soc Mass Spectrom 12, 1296–1301 (2001). https://doi.org/10.1016/S1044-0305(01)00317-8

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  • DOI: https://doi.org/10.1016/S1044-0305(01)00317-8

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