Comparative study of the conformational lock, dissociative thermal inactivation and stability of euphorbia latex and lentil seedling amine oxidases.
The thermal stability of copper/quinone containing amine oxidases from Euphorbia characias latex (ELAO) and lentil seedlings (LSAO) was measured in 100 mM potassium phosphate buffer (pH 7.0) following changes in absorbance at 292 nm. ELAO was shown to be about 10 degrees C more stable than LSAO. The dissociative thermal inactivation of ELAO was studied using putrescine as substrate at different temperatures in the range 47-70 degrees C, and a "conformational lock" was developed using the theory pertaining to oligomeric enzyme. Moreover ELAO was shown to be more stable towards denaturants than LSAO, as confirmed by dodecyl trimethylammonium bromide denaturation curves. A comparison of the numbers of contact sites in inter-subunits of ELAO relative to LSAO led us to conclude that the higher stability of ELAO to temperature and towards denaturants was due to the presence of larger number of contact sites in the conformational lock of the enzyme. This study also gives a putative common mechanism for thermal inactivation of amine oxidases and explains the importance of C-terminal conserved amino acids residues in this class of enzymes.
Amani
M
M
Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran.
Moosavi-Movahedi
A A
AA
Floris
G
G
Longu
S
S
Mura
A
A
Moosavi-Nejad
S Z
SZ
Saboury
A A
AA
Ahmad
F
F
D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D004795:Enzyme Stability; D028481:Euphorbia / Q000737:chemistry*; D006358:Hot Temperature; D007840:Latex / Q000378:metabolism*; D027806:Lens Plant / Q000033:anatomy & histology / Q000201:enzymology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D011489:Protein Denaturation; D011700:Putrescine / Q000378:metabolism; D036226:Seedlings / Q000201:enzymology*; D016415:Sequence Alignment
D007840:Latex; D010940:Plant Proteins; D006631:Amine Oxidase (Copper-Containing); D011700:Putrescine