Antibodies and Fab fragments protect Cu,Zn-SOD against methylglyoxal-induced inactivation

Rukhsana Jabeen, Amin A. Mohammad, Elizabeth C. Elefano, John R. Petersen, Mohammed Saleemuddin

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Methyl glyoxal (MG) is a highly reactive α-oxoaldehyde that plays an important role in non-enzymatic glycosylation reactions, formation of Advanced Glycation End products (AGEs) and other complications associated with hyperglycemia and related disorders. Unlike sugars, glycation by MG is predominantly arginine directed, which is particularly more damaging since arginine residues have a high-frequency occurrence in ligand and substrate recognition sites in receptor and enzyme active sites. Using bovine erythrocyte Cu,Zn-superoxide dismutase (SOD) as model enzyme, the potential of anti-enzyme antibodies in imparting protection against MG-induced inactivation was investigated. A concentration- and time-dependent inactivation of SOD was observed when the enzyme was incubated with MG. The enzyme lost over 80% activity on incubation with 5 mM MG for 5 days. More marked inactivation was observed in 24 h when the MG concentration was raised up to 30 mM. The SOD inactivation was accompanied by the formation of high molecular weight aggregates as revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI/TOF mass spectrometry). Inclusion of specific anti-SOD antibodies raised in rabbits or monomeric Fab fragments derived thereof offered remarkable protection against MG-induced loss in enzyme activity. The protection, however, decreased with increase in the concentration of MG. SELDI/TOF mass spectrometry also revealed that the antibodies restricted the formation of high molecular weight aggregates. The results emphasize the potential of antibody based therapy in combating glycation and related complications.

Original languageEnglish (US)
Pages (from-to)1167-1174
Number of pages8
JournalBiochimica et Biophysica Acta - General Subjects
Volume1760
Issue number8
DOIs
StatePublished - Aug 2006

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Glyoxal
Pyruvaldehyde
Immunoglobulin Fragments
Immunoglobulin Fab Fragments
Superoxide Dismutase
Enzymes
Mass spectrometry
Mass Spectrometry
Antibodies
Arginine
Molecular Weight
Molecular weight
Glycosylation
Advanced Glycosylation End Products
Enzyme activity
Electrophoresis
Sugars
Sodium Dodecyl Sulfate
Hyperglycemia
Ionization

Keywords

  • Glycation
  • Methyl glyoxal
  • Polyclonal antibodies
  • SOD

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Antibodies and Fab fragments protect Cu,Zn-SOD against methylglyoxal-induced inactivation. / Jabeen, Rukhsana; Mohammad, Amin A.; Elefano, Elizabeth C.; Petersen, John R.; Saleemuddin, Mohammed.

In: Biochimica et Biophysica Acta - General Subjects, Vol. 1760, No. 8, 08.2006, p. 1167-1174.

Research output: Contribution to journalArticle

Jabeen, Rukhsana ; Mohammad, Amin A. ; Elefano, Elizabeth C. ; Petersen, John R. ; Saleemuddin, Mohammed. / Antibodies and Fab fragments protect Cu,Zn-SOD against methylglyoxal-induced inactivation. In: Biochimica et Biophysica Acta - General Subjects. 2006 ; Vol. 1760, No. 8. pp. 1167-1174.
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