[Structural-functional study of recombinant forms of onconase].
A method for expression of an onconase gene leading to a soluble form of the protein was developed. The enzymatic and cytotoxic properties of the protein's recombinant forms were studied. Recombinant onconase with an additional N-terminal Met residue isolated in nondenaturing conditions did not substantially differ from the native enzyme in ribonucleolytic activity. The addition of a 33-mer peptide containing auxiliary elements for the simplification of isolation and detection of the recombinant protein did not affect the enzyme properties of onconase. The method proposed is useful for the onconase structure-function relation studies and enables construction of onconase-based fusion proteins for anticancer therapy.
Vorob'ev
I I
II
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, GSP Moscow, 117997 Russia. ivanv@ibch.ru
Ponomarenko
N A
NA
Durova
O M
OM
Kozyr'
A V
AV
Demin
A V
AV
Kolesnikov
A V
AV
Sashchenko
L P
LP
Karpeĭskiĭ
M Ia
MIa
Gabibov
A G
AG
D000818:Animals; D001483:Base Sequence; D004527:Egg Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli; D005260:Female; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010446:Peptide Fragments; D011894:Rana pipiens; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D012260:Ribonucleases / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity
D004527:Egg Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D012260:Ribonucleases; C058703:ranpirnase