Linear dimeric interleukin-2 obtained by the use of a defective herpes simplex viral vector: conformation-activity relationship.
An interleukin-2 dimer, produced enzymatically by a nerve-derived transglutaminase in vitro, is cytotoxic to oligodendrocytes, unlike the immune-derived monomeric interleukin-2. The object of this study was to establish a way to produce a dimer of interleukin-2 in quantities, by means of genetic engineering, and to confirm that the structure of the resulting molecule is critical for its function. A defective herpes simplex virus vector was utilized for overproduction of a dimeric interleukin-2. The resulting linear dimer, which is a translational product, differs from the enzymatically produced dimer, which is a posttranslational modification of interleukin-2. The linear dimer, while retaining the known interleukin-2 activity of monomeric interleukin-2 with respect to mitogenicity on T cells, was not cytotoxic to oligodendrocytes. This finding suggests that the lack of cytotoxicity of the linear dimeric interleukin-2 is not caused by a loss of activity during its preparation but is related to its conformational structure, which evidently does not meet the requirements for cytotoxicity. This study opens the way to the design at the transcriptional level of modified proteins and their efficient production, provided that the new transcript encodes for the desired modification in the protein at the appropriate sites.
Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel.
O
O
Eizenberg
M G
MG
Kaplitt
S
S
Eitan
D W
DW
Pfaff
D L
DL
Hirschberg
M
M
Schwartz
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D017931:DNA Primers; D003673:Defective Viruses; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000737:chemistry* / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000276:immunology; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D018139:Simplexvirus; D012867:Skin / Q000378:metabolism; D013329:Structure-Activity Relationship; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology; D014709:Vero Cells
D017931:DNA Primers; D007376:Interleukin-2; D046911:Macromolecular Substances; D011994:Recombinant Proteins