The pro-sequence facilitates folding of human nerve growth factor from Escherichia coli inclusion bodies

Citation
A. Rattenholl et al., The pro-sequence facilitates folding of human nerve growth factor from Escherichia coli inclusion bodies, EUR J BIOCH, 268(11), 2001, pp. 3296-3303
Citations number
36
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
EUROPEAN JOURNAL OF BIOCHEMISTRY
ISSN journal
0014-2956 → ACNP
Volume
268
Issue
11
Year of publication
2001
Pages
3296 - 3303
Database
ISI
SICI code
0014-2956(200106)268:11<3296:TPFFOH>2.0.ZU;2-B
Abstract
Nerve growth factor (beta -NGF), a neurotrophin required for the developmen t and survival of specific neuronal populations, is translated as a prepro- protein in vivo. While the presequence mediates translocation into the endo plasmic reticulum, the function of the pro-peptide is so far unknown. As th e pro-sequences of several proteins are known to promote folding of the mat ure part, the renaturation behaviour of recombinant human P-NGF pro-protein was compared to that of the mature form. Expression of rh-pro-NGF in Esche richia coil 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 character ization revealed that rh-pro-NGF is a dimer. The pro-peptide could be remov ed by limited proteolysis with trypsin yielding biologically active, mature rh-NGE Furthermore, rh-pro-NGF exhibited biological activity in the same c oncentration range as rh-NGF.