The Protein Journal

, Volume 30, Issue 3, pp 149–158 | Cite as

Glycation of the Muscle-Specific Enolase by Reactive Carbonyls: Effect of Temperature and the Protection Role of Carnosine, Pirydoxamine and Phosphatidylserine

  • Jadwiga Pietkiewicz
  • Agnieszka Bronowicka-Szydełko
  • Katarzyna Dzierzba
  • Regina Danielewicz
  • Andrzej Gamian


Reactive carbonyls such as 4-hydroxy-2-nonenal (4-HNE), trans-2-nonenal (T2 N), acrolein (ACR) can react readily with nucleophilic protein sites forming of advanced glycation end-products (AGE). In this study, the human and pig muscle-specific enolase was used as a protein model for in vitro modification by 4-HNE, T2 N and ACR. While the human enolase interaction with reactive α-oxoaldehyde methylglyoxal (MOG) was demonstrated previously, the effect of 4-HNE, T2 N and ACR has not been identified yet. Altering in catalytic function were observed after the enzyme incubation with these active compounds for 1–24 h at 25, 37 and 45 °C. The inhibition degree of enolase activity occurred in following order: 4-HNE > ACR > MOG > T2 N and inactivation of pig muscle-specific enolase was more effective relatively to human enzyme. The efficiency of AGE formation depends on time and incubation temperature with glycating agent. More amounts of insoluble AGE were formed at 45 °C. We found that pirydoxamine and natural dipeptide carnosine counteracted AGE formation and protected enolase against the total loss of catalytic activity. Moreover, we demonstrated for the first time that phosphatidylserine may significantly protect enolase against decrease of catalytic activity in spite of AGE production.


Muscle-specific enolase Reactive aldehydes Advanced glycation end-products Protective agents 








T2 N



Phosphate buffered saline pH 7.4



This study was supported by Grant No 3564/B/P01/2007/33 from the Ministry of Science and Higher Education and by Wrocław Research Center EIT + under the project “Biotechnologies and advanced medical technologies—BioMed” (POIG.01.01.02-02-003/08-00) financed from the European Regional Development Fund (Operational Programme Innovative Economy, 1.1.2).


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Jadwiga Pietkiewicz
    • 1
  • Agnieszka Bronowicka-Szydełko
    • 1
  • Katarzyna Dzierzba
    • 1
  • Regina Danielewicz
    • 1
  • Andrzej Gamian
    • 1
    • 2
  1. 1.Department of Medical BiochemistryWroclaw Medical UniversityWroclawPoland
  2. 2.Institute of Immunology and Experimental TherapyPolish Academy of SciencesWroclawPoland

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