[Human tumor necrosis factor mutants: preparation and some properties].
Using polymerase chain reaction, a number of mutant genes encoding human tumor necrosis factor (TNF-alpha) with amino acid substitutions and a deletion were obtained. The mutant proteins (muteins) contained point mutations R32H, A33S, F144L, I118M, and I118A; double mutation R32H-F144L; and deletion of four amino acid residues 67-70. The mutant genes were expressed in E. coli under the control of constitutive promoters. A simple purification method for the muteins was developed and their physicochemical properties were studied. All the muteins obtained, except F144L and I118A, were shown by CD and cross-linking to from a spatial structure similar to that of the native TNF-alpha. The collection of muteins was characterized by their biological activity. Mutants R32H and A33S exerted a decreased cytotoxicity against murine fibroblast cell line L929, whereas point mutant F144L and double mutant R32H-F144L were essentially inactive.
Shingarova
L N
LN
National Cancer Institute, Frederick, MD, USA.
Sagaĭdak
L N
LN
Turetskaia
R L
RL
Nedospasov
S A
SA
Esipov
D S
DS
Korobko
V G
VG
D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010957:Plasmids; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D017384:Sequence Deletion; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology*
D017931:DNA Primers; D014409:Tumor Necrosis Factor-alpha
Mutanty faktora nekroza opukholeĭ cheloveka: poluchenie i nekotorye svoĭstva.