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Sequence specific 1H, 13C and 15N resonance assignments of the C-terminal domain of human γS-crystallin

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Abstract

The high solubility and stability of crystallins present in the human eye lens maintains its transparency and refractive index with negligible protein turnover. Monomeric γ-crystallins and oligomeric β-crystallins are made up of highly homologous double Greek key domains. These domains are symmetric and possess higher stability as a result of the complex topology of individual Greek key motifs. γS-crystallin is one of the most abundant structural βγ-crystallins present in the human eye lens. In order to understand the structural stability of individual domains of human γS-crystallin in isolation vis-à-vis full length protein, we set out to structurally characterize its C-terminal domain (abbreviated hereafter as γS-CTD) by solution NMR. In this direction, we have cloned, over-expressed, isolated and purified the γS-CTD. The 2D [15N-1H] HSQC recorded with uniformly 13C/15N labeled γS-CTD showed a highly dispersed spectrum indicating the protein to adopt an ordered conformation. In this paper, we report almost complete sequence-specific 1H, 13C and 15N resonance assignments of γS-CTD using a suite of heteronuclear 3D NMR experiments.

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Abbreviations

γS-CTD:

C-terminal domain of human γS-crystallin

HSQC:

Heteronuclear single quantum correlation

NMR:

Nuclear magnetic resonance

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Acknowledgements

The facilities provided by the National Facility for High Field NMR, supported by the Department of Science and Technology (DST), Department of Biotechnology (DBT), Council of Scientific and Industrial Research (CSIR) and the high field NMR Centre at Indian Institute of Chemical Technology (IICT), Hyderabad are gratefully acknowledged. KVRC acknowledges the financial support of JC Bose fellowship of DST (Govt. of India).

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Author notes

  1. Khandekar Jishan Bari and Shrikant Sharma have share equal authorship.

Authors and Affiliations

  1. Center for Interdisciplinary Sciences, Tata Institute of Fundamental Research, Gopanpally, Hyderabad, 500107, India

    Khandekar Jishan Bari, Shrikant Sharma & Kandala V. R. Chary

  2. Department of Chemical Sciences, Tata Institute of Fundamental Research, 1, Homi Bhabha Road, Colaba, Mumbai, 400005, India

    Kandala V. R. Chary

  3. Indian Institute of Science Education and Research, Berhampur, Odisha, 760010, India

    Kandala V. R. Chary

Authors
  1. Khandekar Jishan Bari
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  2. Shrikant Sharma
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  3. Kandala V. R. Chary
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KJB and SS have contributed equally to this work.

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Correspondence to Kandala V. R. Chary.

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Bari, K.J., Sharma, S. & Chary, K.V.R. Sequence specific 1H, 13C and 15N resonance assignments of the C-terminal domain of human γS-crystallin. Biomol NMR Assign 13, 43–47 (2019). https://doi.org/10.1007/s12104-018-9848-x

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