The unprocessed C-terminal dipeptide of recombinant beta-nerve growth factor determines three stable forms with distinct biological activities.
The processing of polypeptide neurotrophins in the nervous system is poorly understood. In this paper, we provide information on the effects of C-terminal processing of nerve growth factor. Three forms of recombinant mouse beta-nerve growth factor (rNGF) were produced and isolated from insect cells infected with a recombinant baculovirus. The three purified forms of rNGF exhibited distinct biological activities and differed in their abilities to compete with high affinity binding of mouse beta-nerve growth factor (mNGF). However, they were chemically and structurally indistinguishable from each other. All three forms of rNGF differed from mature mNGF from mouse submaxillary gland in that the C-terminal Arg-Gly dipeptide had not been proteolytically removed. Removal of the C-terminal dipeptide by gamma-NGF peptidase treatment converted the three forms into a single form identical with mature mNGF. The above results demonstrate that a single polypeptide of rNGF, due to the presence of a C-terminal dipeptide, exhibits three stable dimeric protein conformations with distinct biological activities. The apparent lack of gamma-NGF peptidase in the nervous system raises the possibility that the biologically significant form of NGF may differ from mature mNGF; such a difference may be of physiological relevance.
Luo
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Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106.
Neet
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D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001667:Binding, Competitive; D015153:Blotting, Western; D002460:Cell Line; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D004151:Dipeptides / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral; D007525:Isoelectric Focusing; D008297:Male; D051379:Mice; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism; D016716:PC12 Cells; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D004151:Dipeptides; D009414:Nerve Growth Factors; D011994:Recombinant Proteins