Thermal dissection of lentil seedling amine oxidase domains by differential scanning calorimetry.
The relationships between the structural and energetic domains of lentil seedling amine oxidase (LSAO) were investigated using modifiers that target the active site and the carbohydrate moiety of the enzyme. An irreversible inhibitor, aminoguanidine, specifically modified the active site of the lentil enzyme, whereas sodium metaperiodate cleaves carbohydrate moieties covalently bound to the native enzyme. Differential scanning calorimetry (DSC) measurements were made on the modified LSAOs. Deconvolution of the reversible thermal DSC profiles of the modified enzyme gave three subpeaks (energetic domains), each of which was assigned to one of the three structural domains of the native protein. Our results led us to conclude that deglycosylation of LSAO has no effect on thermal stability, whereas binding of the inhibitor imparts more stability to the enzyme.
Moosavi-Movahedi
Ali Akbar
AA
Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran. moosavi@ibb.ut.ac.ir
Amani
Mojtaba
M
Moosavi-Nejad
Seyedeh Zahra
SZ
Hashemnia
Sedigheh
S
Ahmad
Faizan
F
Floris
Giovanni
G
Mura
Anna
A
Rezaei-Tavirani
Mostafa
M
Hakimelahi
Gholam Hossein
GH
Saboury
Ali Akbar
AA
Yousefi
Reza
R
D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning; D027806:Lens Plant / Q000201:enzymology*; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D036226:Seedlings / Q000201:enzymology*
D006631:Amine Oxidase (Copper-Containing)