High-level expression of a biologically active human interleukin-6 mutein.
We have constructed two different muteins of interleukin-6 (IL-6) which were expressed in Escherichia coli. Both muteins lack the first 22 N-terminal amino acids of native IL-6 and lack one or the other of the two naturally occurring pairs of cysteines at either position 45 and 51 or position 74 and 84 of IL-6. We found that there was a dramatic increase in the level of IL-6 produced from each mutein clone, compared to the level produced by the wild-type IL-6 clone. We also observed that the yield of soluble and properly refolded mutein IL-6 was highest when the cysteines at position 74 and 84 were left intact. The mutein IL-6 with cysteines at position 74 and 84 was as active as wild-type IL-6 and a lower concentration of the mutein IL-6 was required to reach maximal activity, compared to wild-type IL-6. The mutein IL-6 with cysteines at position 45 and 51 had a much reduced biological activity.
Skelly
S M
SM
ImClone Systems Inc., New York, NY 10014.
Tackney
C
C
Hicklin
D
D
Tamkins
T
T
Goldstein
N
N
Waksal
H
H
Dagan
S
S
D000818:Animals; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D051379:Mice; D009154:Mutation; D013329:Structure-Activity Relationship
D015850:Interleukin-6