Skip to main content

Advertisement

SpringerLink
Log in

Study of Papain–Cystatin Interaction by Intensity Fading MALDI-TOF-MS

  • Published:
The Protein Journal Aims and scope Submit manuscript

Abstract

Intensity fading (IF) matrix assisted laser desorption ionization (MALDI) time of flight (TOF) mass spectrometry (MS ) has become an alternative screening approach for the affinity-binding analysis of proteins and peptides with molecular ligands. In this investigation an attempt has been made to study the protein ligand interaction by intensity fading (IF) MALDI-MS using papain and cystatin as model system for protein-ligand interactions. The intensity fading of cystatin was monitored using various concentration of cystatin ranging from (1 to 8.6 μM) in presence of target protein, papain. The results indeed indicate that the intensity of cystatin decreases upon addition of papain. Furthermore, for the first time we have used IF-MALDI-MS for determining the number of binding sites for cystatin on papain by Scatchard analysis.

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

Similar content being viewed by others

Abbreviations

ACN:

Acetonitrile

DTT:

Diththiothreitol

ESI:

Electrospray ionization

IF:

Intensity fading

MALDI:

Matrix assisted laser desorption ionization

MS:

Mass spectrometry

SA:

Sinapinic acid

SAMDI:

Self-assembled monolayer for MALDI

TFA:

Trifluoroacetic acid

TOF:

Time of flight

References

  1. Northrop DB, Simpson FB (1997) Bioorg Med Chem 5:641–644

    Article  CAS  Google Scholar 

  2. Liesener A, Karst U (2005) Anal Bioanal Chem 382:1451–1464

    Article  CAS  Google Scholar 

  3. Kang MJ, Tholey A, Heinzle E (2001) Rapid Commun Mass Spectrom 15:1327–1333

    Article  CAS  Google Scholar 

  4. Maillard LT, Guerineau V, Badet-Denisot MA, Badet B, Laprevote O, Durand P (2006) Rapid Commun Mass Spectrom 20:666–672

    Article  CAS  Google Scholar 

  5. Yanes O, Aviles XF, Roepstorff P, Jørgensen TJD (2006a) J Am Soc Mass Spectrom 18:359–367

    Article  CAS  Google Scholar 

  6. Villanueva J, Yanes O, Querol E, Serrano L, Aviles XF (2003) Anal Chem 75:3385–3395

    Article  CAS  Google Scholar 

  7. Yanes O, Villanueva J, Querol E, Aviles XF (2004) Drug Discov Targ 3:2 (Suppl.)

  8. Otto HH, Schirmeister T (1997) Chem Rev 97:133–172

    Article  CAS  Google Scholar 

  9. Brown WM, Dziegielewska KM (1997) Prot Sci 6:5–12

    Article  CAS  Google Scholar 

  10. Abrahamson M (1994) Methods in Enzymol 244:685–700

    Article  CAS  Google Scholar 

  11. Zeeuwen PL, Ishida-Yamamoto A, van Vlijmen-Willems IM, Cheng T, Bergers M, Lizuka H, Schalkwijk J (2007) J Invest Dermatol 127:120–128

    Article  CAS  Google Scholar 

  12. Kussmann M, Nordhoff E, Rahbek-Nielsen H, Haebel S, Rossel-Larsen M, Jakobsen L, Gobom J, Mirgorodskaya E, Kroll-Kristensen A, Palm L, Roepstorff P (1997) J Mass Spectrom 32:593–601

    Article  CAS  Google Scholar 

  13. Scatchard G (1949) Ann New York Acad Sci 51:660

    Article  CAS  Google Scholar 

  14. Lindahl P, Alriksson E, Jornvall H, Bjork I (1988) Biochem 27:5074–5082

    Article  CAS  Google Scholar 

  15. Yanes O, Nazabal A, Wenzel R, Zenobi R, Aviles XF (2006b) J Proteome Res 5:2711–2719

    Article  CAS  Google Scholar 

  16. Yanes O, Villanueva J, Querol E, Aviles XF (2007) Nat Prot 2:119–130

    Article  CAS  Google Scholar 

  17. Yanes O, Villanueva J, Querol E, Aviles XF (2005) Mol Cell Proteom 4:1602–1613

    Article  CAS  Google Scholar 

  18. Nazabal A, Wenzel RJ, Zenobi R (2006) Anal Chem 78:3562–3570

    Article  CAS  Google Scholar 

Download references

Acknowledgement

The financial support from CSIR India to M.S is greatly acknowledged. Authors thank Dr. S. Sivaram, Director, NCL, Dr. M K Gurjar (Head, OCS and OCT), Dr. M. M. Bhadbhade (Head, CMC), and Dr. K. N. Ganesh (Director, IISER) for their support and encouragement.

Author information

Authors and Affiliations

  1. Division of Biochemical Sciences, National Chemical Laboratory, Dr. Homi Bhabha Road, Pune, India

    M. Shabab & M. Islam Khan

  2. Mass spectrometry and Proteomics Lab, Division of Organic Chemistry, National Chemical Laboratory, Dr. Homi Bhabha Road, Pune, India

    Mahesh J. Kulkarni

Authors
  1. M. Shabab
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mahesh J. Kulkarni
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Islam Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Mahesh J. Kulkarni or M. Islam Khan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shabab, M., Kulkarni, M.J. & Khan, M.I. Study of Papain–Cystatin Interaction by Intensity Fading MALDI-TOF-MS. Protein J 27, 7–12 (2008). https://doi.org/10.1007/s10930-007-9102-3

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10930-007-9102-3

Keywords

Search

Navigation