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Resonance assignments and secondary structure of a phytocystatin from Ananas comosus

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Abstract

A cDNA encoding a cysteine protease inhibitor, cystatin was cloned from pineapple (Ananas comosus L.) stem. This clone was constructed into an expression vector and expressed in E. coli and purified to homogeneous. The recombinant pineapple cystatins (AcCYS) showed effectively inhibitory activity toward cysteine proteases including papain, bromelain, and cathepsin B. In order to unravel its inhibitory action from structural point of view, multidimensional heteronuclear NMR techniques were used to characterize the structure of AcCYS. The full 1H, 15N, and 13C resonance assignments of AcCYS were determined. The secondary structure of AcCYS was identified by using the assigned chemical shift of 1Hα, 13Cα, 13Cβ, and 13CO through the consensus chemical shift index (CSI). The results of CSI analysis suggest 5 β–strands (residues 45–47, 84–91, 94–104, 106–117, and 123–130) and one α–helix (residues 55–73).

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Acknowledgments

This work was supported by research grants from National Science Council, Taiwan, NSC- 95-2113-M-259-005 and 96-2113-M-259-005 (to Chia-Lin Chyan).

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Authors and Affiliations

  1. Department of Chemistry, National Dong Hwa University, Shoufeng, Hualien, 974, Taiwan, ROC

    Deli Irene, Bo-Jiun Chen, Si-Hung Lo & Chia-Lin Chyan

  2. Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, 402, Taiwan, ROC

    Ting-Hang Liu & Jason T.-C. Tzen

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  1. Deli Irene
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  2. Bo-Jiun Chen
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  3. Si-Hung Lo
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  4. Ting-Hang Liu
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  5. Jason T.-C. Tzen
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  6. Chia-Lin Chyan
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Correspondence to Chia-Lin Chyan.

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Irene, D., Chen, BJ., Lo, SH. et al. Resonance assignments and secondary structure of a phytocystatin from Ananas comosus . Biomol NMR Assign 6, 99–101 (2012). https://doi.org/10.1007/s12104-011-9334-1

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  • DOI: https://doi.org/10.1007/s12104-011-9334-1

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