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Modulation of PP2A activity by Jacalin: is it through caveolae and ER chaperones?

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Abstract

Plant lectins have been reported to affect the proliferation of different human cancer cell line probably by binding to the specific carbohydrate moieties. In the present study, Badan labeled single cysteine mutant (present in the caveolin-1 binding motif) of jacalin (rJacalin) was found to penetrate the target membrane, indicating a protein-protein or protein-membrane interaction apart from its primary mode of binding i.e. protein-carbohydrate interaction. Further, Jacalin treatment has resulted in the movement of the GFP-Caveolin-1 predominantly at the cell-cell contact region with much restricted dynamics. Jacalin treatment has resulted in the perinuclear accumulation of PP2A and dissociation of the PHAP1/PP2A complex. PP2A was found to act as a negative regulator of ERK signaling and a significant decrease in the phosphorylation level of MEK and AKT (T308) in A431. In addition, we have also identified several ER resident proteins including molecular chaperones like ORP150, Hsp70, Grp78, BiP of A431 cells, which were bound to the Jacalin-sepharose column. Among various ER chaperones that were identified, ORP150 was found to present on the cell surface of A431 cells.

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Abbreviations

RT:

Room Temperature

IPA:

Immunoprecipitation assay

TIRFM:

Total Internal Reflection Fluorescence Microscopy

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Acknowledgements

The authors thank Dr. G. C. Mishra, Prof. A. Surolia and Dr. M. Islam Khan for valuable comments, and Mr Anil Lotke and Ms Ashwini Atre for technical assistance. N.A, A.R, S.S, and A.S. are recipients of a senior research fellowship from CSIR and UGC. S.P. is currently postdoctoral fellow, college of Pharmacy, University of Houston, Texas. Financial assistance was provided by the Department of Biotechnology, Government of India.

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Authors and Affiliations

  1. National Centre for Cell Science, Ganeshkhind Road, University of Pune Campus, Pune, 411007, India

    Neesar Ahmed, Satyabrata Pany, Aejazur Rahman, Saumya S. Srivastava, Amita Sneh & Musti V. Krishnasastry

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  1. Neesar Ahmed
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  2. Satyabrata Pany
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  3. Aejazur Rahman
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Correspondence to Musti V. Krishnasastry.

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Video SV1

TIRFM video of the HeLa cells expressing GFP-Caveolin-1 before treatment of Jacalin. The normal ‘kiss-and-run’ dynamics of caveolae are observed [14]. (QT 2506 kb)

Video SV2

TIRFM video of a GFP- Caveolin-1 expressing HeLa cell 1 h after treatment with Jacalin (2 µg/ml). Note the cell surface withdrawal of GFP- Caveolin-1. (QT 3246 kb)

Video SV3

TIRFM video of a GFP- Caveolin-1 expressing HeLa cell 2 h after treatment with Jacalin (2 µg/ml). Note the restricted movement of GFP- Caveolin-1. (QT 3190 kb)

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Ahmed, N., Pany, S., Rahman, A. et al. Modulation of PP2A activity by Jacalin: is it through caveolae and ER chaperones?. Glycoconj J 27, 723–734 (2010). https://doi.org/10.1007/s10719-009-9258-5

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  • DOI: https://doi.org/10.1007/s10719-009-9258-5

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