Skip to main content
SpringerLink
Log in

Unfolding Studies of Escherichia coli Maltodextrin Glucosidase Monitored by Fluorescence Spectroscopy

  • Original Paper
  • Published:
Journal of Biological Physics Aims and scope Submit manuscript

Abstract

Equilibrium unfolding of a 69-kDa monomeric Escherichia coli maltodextrin glucosidase (MalZ) was studied using intrinsic and extrinsic fluorescence spectroscopy. The unfolding transition of MalZ followed a three-state process, involving the formation of a stable intermediate state having more exposed hydrophobic surface. It was found that the protein structure can be easily perturbed by low concentration of guanidium hydrochloride (GdnHCl) and, at a GdnHCl concentration of 2 M, MalZ was denatured completely. The active site of the protein also has been proved to be sensitive to a low concentration of GdnHCl since MalZ deactivated at 0.5 M GdnHCl completely. The surface hydrophobicity and ANS-binding site of the protein have been determined to be 150.7 and 0.24, respectively. Perhaps the formation of the stable unfolding intermediate, having higher surface hydrophobicity, may be one of the reasons for aggregation of MalZ and its recognition by chaperonin GroEL during the assisted folding pathway.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Anfinsen, C.B.: Principles that govern the folding of protein chains. Science 181, 223–230 (1973). doi:10.1126/science.181.4096.223

    Article  ADS  Google Scholar 

  2. Ikemura, I., Takagi, H., Inauye, M.: Requirement of pro-sequence for the production of active subtilisin in Escherichia coli. J. Biol. Chem. 262, 7859–7864 (1987)

    Google Scholar 

  3. Silen, J.L., Agard, D.A.: The alpha-lytic protease pro-region does not require a physical linkage to activate the protease domain in vivo. Nature 341, 462–464 (1989). doi:10.1038/341462a0

    Article  ADS  Google Scholar 

  4. Winther, J.R., Sorensen, P.: Propeptide of carboxypeptidase Y provides a chaperone-like function as well as inhibition of the enzymatic activity. Proc. Natl. Acad. Sci. USA 88, 9330–9334 (1991). doi:10.1073/pnas.88.20.9330

    Article  ADS  Google Scholar 

  5. Matthews, C.B.: Pathways of protein folding. Annu. Rev. Biochem. 62, 653–683 (1993). doi:10.1146/annurev.bi.62.070193.003253

    Article  Google Scholar 

  6. Rudon, R.W., Bedows, E.: Assisted protein folding. J. Biol. Chem. 272, 3125–3128 (1997). doi:10.1074/jbc.272.6.3125

    Article  Google Scholar 

  7. Finkelstein, A.V., Ptitsyn, O.B.: Why do globular proteins fit the limited set of folding patterns? Prog. Biophys. Mol. Biol. 50, 171–190 (1987). doi:10.1016/0079-6107(87)90013-7

    Article  Google Scholar 

  8. Tapio, S., Yeh, F., Shuman, H.A., Boos, W.: The malZ gene of Escherichia coli, a member of the maltose regulon, encodes a maltodextrin glucosidase. J. Biol. Chem. 266, 19450–19458 (1991)

    Google Scholar 

  9. Paul, S., Singh, C., Mishra, S., Chaudhuri, T.K.: The 69 kDa Escherichia maltodextrin glucosidase does not get encapsulated underneath GroES during GroEL/GroES assisted folding. FASEB J. 21, 2874–2885 (2007). doi:10.1096/fj.06-7958com

    Article  Google Scholar 

  10. Chaudhuri, T.K., Das, K.P., Sinha, N.K.: Surface hydrophobicity of a low molecular weight basic trypsin subtilisin inhibitor from marine turtle eggwhite. J. Biochem. 113, 729–733 (1993)

    Google Scholar 

  11. Cardamone, M., Puri, N.K.: Spectrofluorimetric assessment of the surface hydrophobicity of proteins. Biochem. J. 282, 589–593 (1992)

    Google Scholar 

  12. Tanksale, A., Ghatge, M., Deshpande, V.: α-Crystallin binds to the aggregation-prone molten-globule state of alkaline protease: Implications for preventing irreversible thermal denaturation. Protein Sci. 11, 1720–1728 (2002). doi:10.1110/ps.0201802

    Article  Google Scholar 

  13. Sanz, J.M., Gimenez-Gallego, G.: A partly folded state of acidic fibroblast growth factor at low pH. Eur. J. Biochem. 246, 328–335 (1997). doi:10.1111/j.1432-1033.1997.00328.x

    Article  Google Scholar 

  14. Kato, A., Nakai, S.: Hydrophobicity determined by a fluorescence probe method and its correlation with surface properties of proteins. Biochim. Biophys. Acta 624, 13–20 (1980)

    Google Scholar 

  15. Matulis, D., Lovrien, R.: 1-Anilino-8-naphthalene sulfonate anion-protein binding depends primarily on ion pair formation. Biophys. J. 74, 422–429 (1998)

    Article  ADS  Google Scholar 

  16. Jones, S., Thornton, J.M.: Analysis of protein-protein interaction sites using surface patches. J. Mol. Biol. 272, 121–132 (1997).

    Article  Google Scholar 

Download references

Acknowledgements

We thank Professor Winfried Boos, Konstanz, Germany for the gift of plasmids encoding His-tagged MalZ. The authors gratefully acknowledge financial support from the Department of Science and Technology, Govt. of India and Ministry of Human Resources and Development (MHRD), Govt. of India.

Author information

Authors and Affiliations

  1. Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110016, India

    Subhankar Paul, Saroj Mishra & Tapan K. Chaudhuri

  2. Department of Biotechnology and Medical Engineering, National Institute of Technology Rourkela, Rourkela, 769008, India

    Subhankar Paul

  3. Department of Chemistry, Bose Institute, 93/1 A.P.C. Road, Kolkata, 700009, India

    Madhuchhanda Kundu & Kali P. Das

Authors
  1. Subhankar Paul
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Madhuchhanda Kundu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kali P. Das
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Saroj Mishra
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tapan K. Chaudhuri
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tapan K. Chaudhuri.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Paul, S., Kundu, M., Das, K.P. et al. Unfolding Studies of Escherichia coli Maltodextrin Glucosidase Monitored by Fluorescence Spectroscopy. J Biol Phys 34, 539–550 (2008). https://doi.org/10.1007/s10867-008-9117-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10867-008-9117-9

Keywords

Search

Navigation