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- Title
The pro-sequence facilitates folding of human nerve growth factor from Escherichia coli inclusion bodies.
- Authors
Rattenholl, A; Lilie, H; Grossmann, A; Stern, A; Schwarz, E; Rudolph, R
- Abstract
Nerve growth factor (beta-NGF), a neurotrophin required for the development and survival of specific neuronal populations, is translated as a prepro-protein in vivo. While the presequence mediates translocation into the endoplasmic reticulum, the function of the pro-peptide is so far unknown. As the pro-sequences of several proteins are known to promote folding of the mature part, the renaturation behaviour of recombinant human beta-NGF pro-protein was compared to that of the mature form. Expression of rh-pro-NGF in Escherichia coli led to the formation of inclusion bodies (IBs). The presence of the covalently attached pro-sequence significantly increased the yield and rate of refolding with concomitant disulfide bond formation when compared to the in vitro refolding of mature NGF (rh-NGF). Physicochemical characterization revealed that rh-pro-NGF is a dimer. The pro-peptide could be removed by limited proteolysis with trypsin yielding biologically active, mature rh-NGF. Furthermore, rh-pro-NGF exhibited biological activity in the same concentration range as rh-NGF.
- Publication
European journal of biochemistry, 2001, Vol 268, Issue 11, p3296
- ISSN
0014-2956
- Publication type
Journal Article
- DOI
10.1046/j.1432-1327.2001.02232.x