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Probing hemoglobin confinement inside submicron silica tubes using synchrotron SAXS and electrochemical response

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Abstract

The configuration of hemoglobin in solution and confined inside silica nanotubes has been studied using synchrotron small angle X-ray scattering and electrochemical activity. Confinement inside submicron tubes of silica aid in preventing protein aggregation, which is vividly observed for unconfined protein in solution. The radius of gyration (R g) and size polydispersity (p) of confined hemoglobin was found to be lower than that in solution. This was also recently demonstrated in case of confined hemoglobin inside layered polymer capsules. The confined hemoglobin displayed a higher thermal stability with R g and p showing negligible changes in the temperature range 25–75 °C. The differences in configuration between the confined and unconfined protein were reflected in their electrochemical activity. Reversible electrochemical response (from cyclic voltammograms) obtained in case of the confined hemoglobin, in contrary to the observance of only a cathodic response for the unconfined protein, gave direct indication of the differences between the residences of the electroactive heme center in a different orientation compared to that in solution state. The confined Hb showed loss of reversibility only at higher temperatures. The electron transfer coefficient (α) and electron transfer rate constant (k s) were also different, providing additional evidence regarding structural differences between the unconfined and confined states of hemoglobin. Thus, absence of any adverse effects due to confinement of proteins inside the inorganic matrices such as silica nanotubes opens up new prospects for utilizing inorganic matrices as protein “encapsulators”, as well as sensors at varying temperatures.

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Acknowledgments

The authors thank INI (IISc) for TEM, Minakshi Sen (MCBL, IISc) for CLSM imaging. The authors acknowledge the support of the International Centre for Theoretical Physics under ICTP-Elettra Users Program for Synchrotron Radiation.

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Authors and Affiliations

  1. Solid State and Structural Chemistry Unit, Indian Institute of Science, Bangalore, 560012, India

    Soumit S. Mandal, Brindha Nagarajan & Aninda J. Bhattacharyya

  2. Institute of Biophysics and Nanosystems Research, Austrian Academy of Sciences, Schmiedlstrasse 6, 8042, Graz, Austria

    H. Amenitsch

Authors
  1. Soumit S. Mandal
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  2. Brindha Nagarajan
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  3. H. Amenitsch
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  4. Aninda J. Bhattacharyya
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Corresponding author

Correspondence to Aninda J. Bhattacharyya.

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Mandal, S.S., Nagarajan, B., Amenitsch, H. et al. Probing hemoglobin confinement inside submicron silica tubes using synchrotron SAXS and electrochemical response. Eur Biophys J 42, 371–382 (2013). https://doi.org/10.1007/s00249-013-0886-0

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  • DOI: https://doi.org/10.1007/s00249-013-0886-0

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