scholarly journals Establishing isostructural metal substitution in metalloproteins using1H NMR, circular dichroism, and Fourier transform infrared spectroscopy

1995 ◽  
Vol 4 (8) ◽  
pp. 1571-1576 ◽  
Author(s):  
Dean L. Pountney ◽  
Colin J. Henehan ◽  
Milan Vasak
Keyword(s):  
Fourier Transform ◽  
Infrared Spectroscopy ◽  
Circular Dichroism ◽  
Fourier Transform Infrared Spectroscopy ◽  
Fourier Transform Infrared ◽  
Metal Substitution ◽  
Circular Dichroïsm

Effect of oxidative stress on stability and structure of neurofilament proteins

2000 ◽  
Vol 78 (6) ◽  
pp. 667-674 ◽  
Author(s):  
Sylvie Gélinas ◽  
Camille Chapados ◽  
Marc Beauregard ◽  
Isabelle Gosselin ◽  
Maria-Grazia Martinoli
Keyword(s):  
Oxidative Stress ◽  
Fourier Transform ◽  
Infrared Spectroscopy ◽  
Circular Dichroism ◽  
Free Radical ◽  
Fourier Transform Infrared Spectroscopy ◽  
Fourier Transform Infrared ◽  
Random Coil ◽  
Neurofilament Proteins ◽  
Circular Dichroïsm

Neurofilament proteins are highly phosphorylated molecules in the axonal compartment of the adult nervous system. We report the structural analysis of neurofilament proteins after oxidative damage. SDS-PAGE, immunoblotting, circular dichroism, and Fourier transform infrared spectroscopy were used to investigate the relative sensitivity of neurofilaments to oxidative stress and to identify changes in their molecular organization. An ascorbate-Fe+3-O2 buffer system as well as catechols were used to generate free radicals on a substrate of phosphorylated and dephosphorylated neurofilaments. By Fourier Transform Infrared spectroscopy and circular dichroism, we established that the neurofilament secondary structure is mainly composed of α-helices and that after free radical damage of the peptide backbone of neurofilaments, those helices are partly modified into β-sheet and random coil structures. These characteristic reorganizations of the neurofilament structure after oxidative exposure suggest that free radical activity might play an important role in the biogenesis of the cytoplasmic inclusions found in several neurodegenerative diseases.Key words: neurofilaments, oxidative stress, neurodegeneration, phosphorylation, infrared spectroscopy, circular dichroism.

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