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Conformational Transitions in Ariesaema curvatum Lectin: Characterization of an Acid Induced Active Molten Globule

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Abstract

Biophysical characterization of a lectin from Ariesaema curvatum (ACL) was carried out using steady state as well as time resolved fluorescence and CD spectroscopy under various denaturing conditions. An intermediate with altered tryptophan microenvironment was detected in the phase diagram, which exibited pronounced secondary structure and hemagglutinating activity in presence of 0.25 M Gdn–HCl. An acid induced molten- globule like structure possessing activity and higher thermostability was detected. Transition to the molten globule state was reversible in nature. The lectin retained hemagglutinating activity even after incubation at 95 °C. Both chemical and thermal unfolding of the lectin were found to consist of multistate processes. Fluorescence quenching of ACL was strong with acrylamide and KI. The single tryptophan was found to be surrounded by high density of the positively charged amino acid residues as shown by a ten fold higher Ksv for KI compared to that for CsCl. The average lifetime of tryptophan fluorescence increased from 1.24 ns in the native state to 1.72 ns in the denatured state.

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Acknowledgments

The authors wish to thank Dr. Arvind Sahu, NCCS, Pune, and Prof. G. Krishnamoorthy, TIFR, Mumbai for permitting to use the CD spectrometer and time-resolved fluorescence facility, respectively. Urvashi thanks UGC, Govt. of India, for a research fellowship.

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Authors and Affiliations

  1. Division of Biochemical Sciences, National Chemical Laboratory, Pune, Maharashtra, 411008, India

    Urvashi Sharma, Sushama M. Gaikwad & C. G. Suresh

  2. Department of Molecular Biology and Biochemistry, Guru Nanak Dev University, Amritsar, 143005, Punjab, India

    Vikram Dhuna, Jatinder Singh & Sukhdev Singh Kamboj

Authors
  1. Urvashi Sharma
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  2. Sushama M. Gaikwad
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  3. C. G. Suresh
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  4. Vikram Dhuna
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  5. Jatinder Singh
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  6. Sukhdev Singh Kamboj
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Correspondence to Sushama M. Gaikwad.

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Sharma, U., Gaikwad, S.M., Suresh, C.G. et al. Conformational Transitions in Ariesaema curvatum Lectin: Characterization of an Acid Induced Active Molten Globule. J Fluoresc 21, 753–763 (2011). https://doi.org/10.1007/s10895-010-0766-2

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  • DOI: https://doi.org/10.1007/s10895-010-0766-2

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