Structure and stability of recombinant protein depend on the extra N-terminal methionine residue: S6 permutein from direct and fusion expression systems.
Two permuted variants of S6 ribosomal protein were obtained in direct and fusion expression systems, respectively. The product of direct expression contained the extra N-terminal methionine residue. The structural properties and conformational stability of these permuteins were compared using 1-D (1)H-NMR, circular dichroism, intrinsic fluorescence, differential scanning calorimetry and resistance to urea-induced unfolding. A pronounced difference in all the parameters studied has been demonstrated. This means that the structure of recombinant protein can be sensitive to peculiarities of the expression and purification procedures, leading particularly to the presence or absence of the Met at the first position in the target protein sequence.
Uversky
V N
VN
Institute for Biological Instrumentation, Russian Academy of Sciences, 142292, Pushchino, Moscow Region, Russia.
Abdullaev
Z K
ZK
Arseniev
A S
AS
Bocharov
E V
EV
Dolgikh
D A
DA
Latypov
R F
RF
Melnik
T N
TN
Vassilenko
K S
KS
Kirpichnikov
M P
MP
D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D009682:Magnetic Resonance Spectroscopy; D008715:Methionine / Q000737:chemistry; D011487:Protein Conformation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics*; D016963:Thermus thermophilus / Q000235:genetics; D014364:Tryptophan / Q000737:chemistry; D014508:Urea
D011994:Recombinant Proteins; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D014364:Tryptophan; D014508:Urea; D008715:Methionine