A non-native alpha-helix is formed in the beta-sheet region of the molten globule state of canine milk lysozyme.
The native and the molten globule states (N and MG states, respectively) of canine milk lysozyme (CML) were examined by CD spectroscopy and AGADIR algorithm, a helix-coil transition program. It revealed that the helical content of the MG state was higher than that of the N-state, suggesting that non-native alpha-helix is formed in the MG state of CML. Using AGADIR, it indicated the possibility of alpha-helix formation in the third beta-strand region in the MG state. To investigate this possibility, we designed a mutant, Q58P, in which the helical propensity of the MG state was significantly decreased around the third beta-strand region. It appeared that the absolute ellipticity value at 222 nm of the mutant in the MG state was smaller than that of the wild-type protein. It could be assumed that the non-native alpha-helix is formed around the third beta-strand region of wild-type CML in the MG state.
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
Masahiro
M
Watanabe
Yoshihiro
Y
Kobashigawa
Tomoyasu
T
Aizawa
Makoto
M
Demura
Katsutoshi
K
Nitta
D000465:Algorithms; D019943:Amino Acid Substitution / Q000235:genetics*; D000818:Animals; D002942:Circular Dichroism; D004285:Dogs; D008892:Milk / Q000201:enzymology*; D009113:Muramidase / Q000737:chemistry* / Q000235:genetics; D017354:Point Mutation / Q000235:genetics*; D011489:Protein Denaturation / Q000235:genetics; D017510:Protein Folding; D017433:Protein Structure, Secondary
D009113:Muramidase