Enhanced proliferative effects of a baculovirus-produced fusion protein of insulin-like growth factor and alpha(1)-proteinase inhibitor and improved anti-elastase activity of the inhibitor with glutamate at position 351.
Alpha(1)-proteinase inhibitor (API) was coupled at the C-terminus of a human insulin-like growth factor (IGF) analog to facilitate its production in insect cells. This fusion protein significantly increased thymidine incorporation into HL-60 cells as compared with the incorporation observed with an equivalent molar mixture of the IGF analog and API. The M351E variant of API has been previously shown to reduce aggregate formation in prokaryotic expression systems. When the oxidation-sensitive methionine 351 of the inhibitor was changed to glutamate, the M351E variant was secreted in larger amounts from insect cells than the corresponding fusion protein with wild-type API. The M351E fusion protein and the corresponding chimera containing the wild-type API were tested for their capacity to inhibit human neutrophil elastase. The M351E variant was a more potent elastase inhibitor than the fusion protein containing the wild-type analog, whereas the proliferative activity of both chimeras was identical. The described mitogenic effect of the chimera and the improved anti-elastase activity of the M351E variant are two ideal properties for therapeutic agents acting in pathological situations where cell proliferation and inhibition of neutrophil elastase have to take place simultaneously, such as during wound healing.
Sandoval
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C
Endocrine Laboratory, McGill University Health Centre, 687 avenue des pins, ouest, Montreal, Quebec, Canada H3A 1A1.
Curtis
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H
Congote
Luis F
LF
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002455:Cell Division; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000235:genetics; D018698:Glutamic Acid / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D007313:Insecta; D007335:Insulin-Like Growth Factor II / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D009479:Neuropeptides; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors; D017354:Point Mutation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013936:Thymidine / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology
D009479:Neuropeptides; D011993:Recombinant Fusion Proteins; D000515:alpha 1-Antitrypsin; C075028:bombyxins; D018698:Glutamic Acid; D007335:Insulin-Like Growth Factor II; D008715:Methionine; D010196:Pancreatic Elastase; D013936:Thymidine