Pro-sequence assisted folding and disulfide bond formation of human nerve growth factor††Dedicated to Rita Levi-Montalcini.22Edited by R. Huber

Rattenholl, Anke; Ruoppolo, Margherita; Flagiello, Angela; Monti, Maria; Vinci, Floriana; Marino, Gennaro; Lilie, Hauke; Schwarz, Elisabeth; Rudolph, Rainer

doi:10.1006/jmbi.2000.4295

Cited by 89 publications

(86 citation statements)

References 33 publications

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“…In case of NGF, the pro-form has recently been attributed pro-apoptotic functions (13,15). In addition, we have previously demonstrated that the pro-peptide significantly contributes to in vitro oxidative folding (10,11). The original objective of the presented work was to show a similar role for the pro-peptide of BMP-2.…”

Section: The Mature Part Of Bmp-2 Influences the Spectroscopic Propermentioning

confidence: 87%

Biophysical Comparison of BMP-2, ProBMP-2, and the Free Pro-peptide Reveals Stabilization of the Pro-peptide by the Mature Growth Factor

Hillger

Herr²,

Rudolph

et al. 2005

Journal of Biological Chemistry

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Pro-forms of growth factors have received intensive scientific attention recently because in some cases different biological activities have been ascribed compared with the mature growth factors. Examples are the pro-apoptotic role of the nerve growth factor (NGF) proform (proNGF) or the latency of the transforming growth factor (TGF)-␤ pro-form (proTGF-␤). To investigate a possible biological function of the pro-form of bone morphogenetic protein (BMP)-2, a member of the TGF-␤ family, mature BMP-2, proBMP-2, and the isolated pro-peptide were recombinantly produced in Escherichia coli cells, and a biophysical comparison was performed. Protocols were developed that allowed efficient refolding and subsequent purification of the proteins. ProBMP-2 could be processed to an N-terminally truncated form of BMP-2, digit removed BMP-2 (drBMP-2), that possessed biological activity, i.e. it induced ectopic bone formation. Bone inducing activity was also displayed by proBMP-2. The three proteins were characterized both by fluorescence and CD spectroscopy. From these analyses, predominant ␤-sheet secondary structural elements in the pro-peptide were deduced. The thermodynamic stability of the pro-peptide was determined by chemical unfolding assays. As in the case of NGF/proNGF, the mature part of BMP-2 stabilized the structure of the pro-peptide moiety. However, in contrast to NGF/proNGF, the pro-peptide did not stimulate oxidative folding of the mature part in vitro.

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Section: The Mature Part Of Bmp-2 Influences the Spectroscopic Propermentioning

confidence: 87%

Biophysical Comparison of BMP-2, ProBMP-2, and the Free Pro-peptide Reveals Stabilization of the Pro-peptide by the Mature Growth Factor

Hillger

Herr²,

Rudolph

et al. 2005

Journal of Biological Chemistry

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“…We could show that oxidative folding in vitro giving rise to the native homodimer is significantly stimulated by the pro-peptide [7,8]. Thus, the question arises as to whether the pro-peptide exists in isolation in a dimeric form.…”

Section: The Isolated Ngf Pro-peptide Is Monomericmentioning

confidence: 99%

“…The 103 amino acids comprising pro-peptide that does not possess any cystine residues (Fig. 1C) is known to promote correct maturation in vivo and guides oxidative refolding of the mature moiety in vitro [5][6][7][8]. Thus, proNGF joins the group of those proteins, of which structure formation is significantly aided by their propeptides that do not engage in redox reactions (for review, see [9,10]).…”

Section: Introductionmentioning

confidence: 99%

The mature part of proNGF induces the structure of its pro-peptide

KLIEMANNEL

2004

FEBS Letters

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Human nerve growth factor (NGF) belongs to the structural family of cystine knot proteins, characterized by a disulfide pattern in which one disulfide bond threads through a ring formed by a pair of two other disulfides connecting two adjacent b-strands. Oxidative folding of NGF revealed that the pro-peptide of NGF stimulates in vitro structure formation. In order to learn more about this folding assisting protein fragment, a biophysical analysis of the pro-peptide structure has been performed. While proNGF is a non-covalent homodimer, the isolated pro-peptide is monomeric. No tertiary contacts stabilize the pro-peptide in its isolated form. In contrast, the pro-peptide appears to be structured when bound to the mature part. The results presented here demonstrate that the mature part stabilizes the structure in the pro-peptide region. This is the first report that provides a biophysical analysis of a pro-peptide of the cystine knot protein family.

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“…In vivo, they are translated as a pre-proprotein which is proteolytically processed at a conserved arginine motif to yield the mature form. The presequence mediates the translocation of the proteins to the endoplasmic reticulum, and the propeptide region has been shown to promote folding of the mature region (Rattenholl et al 2001a). Recent in vitro experiments have shown that a dimeric recombinant human NGF-pro-protein maintains biological activity (Rattenholl et al 2001b), but less is known about the general function of the preproregions in vivo.…”

Section: Introductionmentioning

confidence: 99%

Molecular Evolution of Three Avian Neurotrophin Genes: Implications for Proregion Functional Constraints

Sehgal

Lovette

2003

Journal of Molecular Evolution

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Abstract. Neurotrophin proteins are essential for the survival, differentiation, and maintenance of neurons in the peripheral and central nervous systems. Recent studies have shown that the unprocessed proforms of the neurotrophins are preferential high-affinity ligands for p75 NTR and potent inducers of p75 NTR -mediated cell death. Here, we explore differences in the selective constraints acting on the proregions of the three avian neurotrophin genes-NT-3, BDNF, and NGF-in an explicit phylogenetic context. We found a 50-fold difference in levels of constraint as estimated by d N /d S ratios, with the NGF proregion showing the lowest degree of constraint and BDNF the highest. These patterns suggest that the high conservation exhibited by the BDNF proregion results from intense functional constraints that are relaxed in NGF and somewhat relaxed in NT-3. The proregion of BDNF is likely to have a function that differentiates it from the corresponding regions of the NGF and NT-3 genes, suggesting that BDNF is the avian neurotrophin most likely to be used both in its precursor and mature forms in vivo.

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Pro-sequence assisted folding and disulfide bond formation of human nerve growth factor††Dedicated to Rita Levi-Montalcini.22Edited by R. Huber

Cited by 89 publications

References 33 publications

Biophysical Comparison of BMP-2, ProBMP-2, and the Free Pro-peptide Reveals Stabilization of the Pro-peptide by the Mature Growth Factor

Biophysical Comparison of BMP-2, ProBMP-2, and the Free Pro-peptide Reveals Stabilization of the Pro-peptide by the Mature Growth Factor

The mature part of proNGF induces the structure of its pro-peptide

Molecular Evolution of Three Avian Neurotrophin Genes: Implications for Proregion Functional Constraints

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