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Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues
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| Zeitschriftentitel: | European Journal of Biochemistry |
|---|---|
| Personen und Körperschaften: | , , , , , , , , , , , , |
| In: | European Journal of Biochemistry, 204, 1992, 2, S. 649-655 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
|
| Schlagwörter: |
| author_facet |
CACCIA, Paolo NITTI, Gianpaolo CLETINI, Ornella PUCCI, Piero RUOPPOLO, Margherita BERTOLERO, Federico VALSASINA, Barbara ROLETTO, Fulvia CRISTIANI, Cinzia CAUET, Gilles SARMIENTOS, Paolo MALORNI, Antonio MARINO, Gennaro CACCIA, Paolo NITTI, Gianpaolo CLETINI, Ornella PUCCI, Piero RUOPPOLO, Margherita BERTOLERO, Federico VALSASINA, Barbara ROLETTO, Fulvia CRISTIANI, Cinzia CAUET, Gilles SARMIENTOS, Paolo MALORNI, Antonio MARINO, Gennaro |
|---|---|
| author |
CACCIA, Paolo NITTI, Gianpaolo CLETINI, Ornella PUCCI, Piero RUOPPOLO, Margherita BERTOLERO, Federico VALSASINA, Barbara ROLETTO, Fulvia CRISTIANI, Cinzia CAUET, Gilles SARMIENTOS, Paolo MALORNI, Antonio MARINO, Gennaro |
| spellingShingle |
CACCIA, Paolo NITTI, Gianpaolo CLETINI, Ornella PUCCI, Piero RUOPPOLO, Margherita BERTOLERO, Federico VALSASINA, Barbara ROLETTO, Fulvia CRISTIANI, Cinzia CAUET, Gilles SARMIENTOS, Paolo MALORNI, Antonio MARINO, Gennaro European Journal of Biochemistry Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues Biochemistry |
| author_sort |
caccia, paolo |
| spelling |
CACCIA, Paolo NITTI, Gianpaolo CLETINI, Ornella PUCCI, Piero RUOPPOLO, Margherita BERTOLERO, Federico VALSASINA, Barbara ROLETTO, Fulvia CRISTIANI, Cinzia CAUET, Gilles SARMIENTOS, Paolo MALORNI, Antonio MARINO, Gennaro 0014-2956 1432-1033 Wiley Biochemistry http://dx.doi.org/10.1111/j.1432-1033.1992.tb16678.x <jats:p>The production of recombinant human basic fibroblast growth factor (rhbFGF) in <jats:italic>Escherichia coli</jats:italic> cells yielded active forms of this polypeptide which, however, displayed a high degree of instability towards oxidative processes. Biochemical studies in our laboratory and those of others indicated that the reactivity of the four cysteine residues was the main cause of the observed instability. Several attempts to obtain more stable derivatives of rhbFGF were carried out by modification of the sulfhydryl groups. Among these, treatment of rhbFGF with iodoacetic acid led to the isolation of a partially carboxymethylated form (Cm‐FGF). Peptide mapping analysis of the modified protein showed that two cysteines (78 and 96) were blocked by a carboxymethyl group. The remaining cysteines (34 and 101) were not modified under the conditions used and were found to be in the reduced form. Cm‐FGF and unmodified rhbFGF showed similar affinity both for heparin and for high‐affinity receptors. Cm‐FGF was more stable than the unmodified molecule as measured by HPLC and SDS/PAGE analysis. Interestingly, Cm‐FGF was more active than unmodified rhbFGF in stimulating proliferation of endothelial cells and DNA synthesis in 3T3 fibroblasts. This new derivative could represent a desirable complementation to rhbFGF for the development of more stable pharmaceutical formulations in wound healing applications.</jats:p> Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues European Journal of Biochemistry |
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10.1111/j.1432-1033.1992.tb16678.x |
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1992 |
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European Journal of Biochemistry |
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49 |
| title |
Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_unstemmed |
Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_full |
Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_fullStr |
Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_full_unstemmed |
Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_short |
Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_sort |
stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| topic |
Biochemistry |
| url |
http://dx.doi.org/10.1111/j.1432-1033.1992.tb16678.x |
| publishDate |
1992 |
| physical |
649-655 |
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<jats:p>The production of recombinant human basic fibroblast growth factor (rhbFGF) in <jats:italic>Escherichia coli</jats:italic> cells yielded active forms of this polypeptide which, however, displayed a high degree of instability towards oxidative processes. Biochemical studies in our laboratory and those of others indicated that the reactivity of the four cysteine residues was the main cause of the observed instability. Several attempts to obtain more stable derivatives of rhbFGF were carried out by modification of the sulfhydryl groups. Among these, treatment of rhbFGF with iodoacetic acid led to the isolation of a partially carboxymethylated form (Cm‐FGF). Peptide mapping analysis of the modified protein showed that two cysteines (78 and 96) were blocked by a carboxymethyl group. The remaining cysteines (34 and 101) were not modified under the conditions used and were found to be in the reduced form. Cm‐FGF and unmodified rhbFGF showed similar affinity both for heparin and for high‐affinity receptors. Cm‐FGF was more stable than the unmodified molecule as measured by HPLC and SDS/PAGE analysis. Interestingly, Cm‐FGF was more active than unmodified rhbFGF in stimulating proliferation of endothelial cells and DNA synthesis in 3T3 fibroblasts. This new derivative could represent a desirable complementation to rhbFGF for the development of more stable pharmaceutical formulations in wound healing applications.</jats:p> |
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| author | CACCIA, Paolo, NITTI, Gianpaolo, CLETINI, Ornella, PUCCI, Piero, RUOPPOLO, Margherita, BERTOLERO, Federico, VALSASINA, Barbara, ROLETTO, Fulvia, CRISTIANI, Cinzia, CAUET, Gilles, SARMIENTOS, Paolo, MALORNI, Antonio, MARINO, Gennaro |
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| description | <jats:p>The production of recombinant human basic fibroblast growth factor (rhbFGF) in <jats:italic>Escherichia coli</jats:italic> cells yielded active forms of this polypeptide which, however, displayed a high degree of instability towards oxidative processes. Biochemical studies in our laboratory and those of others indicated that the reactivity of the four cysteine residues was the main cause of the observed instability. Several attempts to obtain more stable derivatives of rhbFGF were carried out by modification of the sulfhydryl groups. Among these, treatment of rhbFGF with iodoacetic acid led to the isolation of a partially carboxymethylated form (Cm‐FGF). Peptide mapping analysis of the modified protein showed that two cysteines (78 and 96) were blocked by a carboxymethyl group. The remaining cysteines (34 and 101) were not modified under the conditions used and were found to be in the reduced form. Cm‐FGF and unmodified rhbFGF showed similar affinity both for heparin and for high‐affinity receptors. Cm‐FGF was more stable than the unmodified molecule as measured by HPLC and SDS/PAGE analysis. Interestingly, Cm‐FGF was more active than unmodified rhbFGF in stimulating proliferation of endothelial cells and DNA synthesis in 3T3 fibroblasts. This new derivative could represent a desirable complementation to rhbFGF for the development of more stable pharmaceutical formulations in wound healing applications.</jats:p> |
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| imprint_str_mv | Wiley, 1992 |
| institution | DE-D275, DE-Bn3, DE-Brt1, DE-D161, DE-Zwi2, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229 |
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| spelling | CACCIA, Paolo NITTI, Gianpaolo CLETINI, Ornella PUCCI, Piero RUOPPOLO, Margherita BERTOLERO, Federico VALSASINA, Barbara ROLETTO, Fulvia CRISTIANI, Cinzia CAUET, Gilles SARMIENTOS, Paolo MALORNI, Antonio MARINO, Gennaro 0014-2956 1432-1033 Wiley Biochemistry http://dx.doi.org/10.1111/j.1432-1033.1992.tb16678.x <jats:p>The production of recombinant human basic fibroblast growth factor (rhbFGF) in <jats:italic>Escherichia coli</jats:italic> cells yielded active forms of this polypeptide which, however, displayed a high degree of instability towards oxidative processes. Biochemical studies in our laboratory and those of others indicated that the reactivity of the four cysteine residues was the main cause of the observed instability. Several attempts to obtain more stable derivatives of rhbFGF were carried out by modification of the sulfhydryl groups. Among these, treatment of rhbFGF with iodoacetic acid led to the isolation of a partially carboxymethylated form (Cm‐FGF). Peptide mapping analysis of the modified protein showed that two cysteines (78 and 96) were blocked by a carboxymethyl group. The remaining cysteines (34 and 101) were not modified under the conditions used and were found to be in the reduced form. Cm‐FGF and unmodified rhbFGF showed similar affinity both for heparin and for high‐affinity receptors. Cm‐FGF was more stable than the unmodified molecule as measured by HPLC and SDS/PAGE analysis. Interestingly, Cm‐FGF was more active than unmodified rhbFGF in stimulating proliferation of endothelial cells and DNA synthesis in 3T3 fibroblasts. This new derivative could represent a desirable complementation to rhbFGF for the development of more stable pharmaceutical formulations in wound healing applications.</jats:p> Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues European Journal of Biochemistry |
| spellingShingle | CACCIA, Paolo, NITTI, Gianpaolo, CLETINI, Ornella, PUCCI, Piero, RUOPPOLO, Margherita, BERTOLERO, Federico, VALSASINA, Barbara, ROLETTO, Fulvia, CRISTIANI, Cinzia, CAUET, Gilles, SARMIENTOS, Paolo, MALORNI, Antonio, MARINO, Gennaro, European Journal of Biochemistry, Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues, Biochemistry |
| title | Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_full | Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_fullStr | Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_full_unstemmed | Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_short | Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_sort | stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| title_unstemmed | Stabilization of recombinant human basic fibroblast growth factor by chemical modifications of cysteine residues |
| topic | Biochemistry |
| url | http://dx.doi.org/10.1111/j.1432-1033.1992.tb16678.x |