- Email: [email protected]
Val-Geninthiocin: A thiopeptide antibiotic produced by Streptomyces sp. RSF18
S76
Abstracts / Journal of Biotechnology 136S (2008) S75–S98
I3-O-008
I3-O-012
Val-Geninthiocin: A thiopeptide antibiotic produced by Streptomyces sp. RSF18
Investigating the action mechanism for a natural active compound honokiol by quantitative proteomics
Imran Sajid 1,2,∗ , Khaled A. Shaaban 1 , Holm Frauendorf 1 , Shahida Hasnain 2 , Hartmut Laatsch 1
Shufang Liang 1,∗ , Yuhuan Xu 1 , Bo Ling 2 , Xia Zhao 2 , Jin Zhou 1 , Lijuan Chen 1 , Yuquan Wei 1
1
Institute of Organic and Biomolecular Chemistry, Tammanstrasse 2, Georg-August University of Gottingen 37077, Germany 2 Department of Microbiology and Molecular Genetics, University of the Punjab, Quaid-e-Azam Campus, Lahore 54590, Pakistan
1
Thiopeptide antibiotics are forming a group of cyclic peptides characterized by several common structural features, such as oxazole and thiazole units and unusual amino acids; especially dehydroamino acids are typical (Chiu et al., 1999). Thiopeptides have been used as antibacterial agents against Gram-positive bacteria and anaerobes, including pathogens resistant to antibiotics currently in use (Nagai et al., 2003), and also have potential as growth inhibitors of the human malaria parasite (Rogers et al., 1998). ValGeninthiocin (1), a new member of thiopeptide antibiotics, was isolated from the mycelium of Streptomyces sp. RSF18, along with the closely related geninthiocin (2) and the macrolide, chalcomycin. Compound 1 showed potent activity against Gram-positive bacteria and minor antifungal activity, while it was not effective against Gram-negative bacteria or microoalgae. By intensive NMR and MS studies, structure of 1 was established as a thiopeptide containing several oxazole and thiazole units and a number of unusual amino acids. We are describing here the fermentation, isolation and structure elucidation as well as the biological activity of 1.
E-mail address: [email protected] (S. Liang).
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China 2 West China Second Hospital, Sichuan University, Chengdu 610041, China
Chiu, M.L., Folche, M., Katoh, T., Puglia, A.M., Vohradsky, J., Yun, B.S., Seto, H., Thompson, C.J., 1999. Broad spectrum thiopeptide recognition specificity of the Streptomyces lividans TipAL protein and its role in regulating gene expression. J. Biol. Chem. 274, 20578–20586. Nagai, K., Kamigiri, K., Arao, N., Suzummura, K., Kawano, Y., Yamaoka, M., Zhang, H., Watanabe, M., Zuzki, K., 2003. YM-266183 and YM-266184, novel thiopeptide antibiotics produced by Bacillus cereus isolated from a Marine Sponge. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological properties. J. Antibiot. 56, 123–128. Rogers, M.J., Cundliffe, E.T., McCutchan, F., 1998. The antibiotic micrococcin is a potent inhibitor of growth and protein synthesis in the malaria parasite. Antimicrob. Agents Chemother. 42, 715–716.
The quantitative proteomics brings about a new method to investigate the action mechanism of natural active compounds from organisms. The SILAC (stable-isotope labelling by amino acids in cell culture) combined with mass spectrometry (MS) has emerged as a simple and powerful quantitative proteomic technique (Ong et al., 2002). Honokiol (HNK), an active component purified from Magnolia officinalis, a plant used in traditional Chinese, exhibits antitumor effects by inhibiting tumor growth (Bai et al., 2003), while proteins involved in antitumor activity in proteomic level are still unclear. In our study, HNK could inhibit cell proliferation and induce apoptosis of HeLa and HepG2 cells in a concentration- and time-dependent manner. We applied the SILAC–MS technique to analyze the differential proteome profiling of cells treated by HNK to investigate key proteins responsible for HNK activities. The changed proteins covered a broad variety of cellular functions including metabolism, signal transduction etc., which indicated HNK performs cytotoxicity to tumor cells through co-operating of many proteins and different pathways. Among these changed proteins, IQGAP1, -tubin, peroxiredoxin-6 and HSP70 etc. proteins were down-regulated significantly, while proteins including annexin A2 etc. were upregulated after HNK treatment. Since IQGAP1 plays important roles in cell adhesion and migration (Noritake et al., 2005), we supposed that HNK may have effects on cell migration through IQGAP1 based on our MS datasheet. Our further scratch migration assay showed that the migration inhibition of HepG2 cells can be induced by HNK, the RNA and protein expression level of IQGAP1 were respectively decreased obviously in HepG2 cells exposed to 10 g/ml HNK for 24 h. Therefore, the down-regulated expression of IQGAP1 by HNK treatment was correlated with cell migration, and HNK probably inhibits cell proliferation and migration through IQGAP1 expression changes and its interactions with other proteins.
doi:10.1016/j.jbiotec.2008.07.169
References
Keywords: Val-Geninthiocin; Thiopeptide antibiotic; Streptomyces References
Bai, X., Cerimele, F., Ushio-Fukai, M., Waqas, M., Campbell, P.M., Govindarajan, B., Der, C.J., Battle, T., Frank, D.A., Ye, K., Murad, E., Dubiel, W., Soff, G., Arbieser, J.L., 2003. Honokiol, a small molecular weight natural product, inhibits angiogenesis in vitro and tumor growth in vivo. J. Biol. Chem. 278, 35501–35507. Noritake, J., Watanabe, T., Sato, K., Wang, S., Kaibuchi, K., 2005. IQGAP1: a key regulator of adhesion and migration. J. Cell Sci. 118, 2085–2092. Ong, S.E., Blagoev, B., Kratchmarova, I., Kristensen, D.B., Steen, H., Pandey, A., Mann, M., 2002. Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics. Mol. Cell. Proteomics 1, 376–386.
doi:10.1016/j.jbiotec.2008.07.170
∗ Corresponding author at: Institute of Organic and Biomolecular Chemistry, Tammanstrasse 2, Georg-August University of Gottingen 37077, Germany.
Abstracts / Journal of Biotechnology 136S (2008) S75–S98
I3-O-008
I3-O-012
Val-Geninthiocin: A thiopeptide antibiotic produced by Streptomyces sp. RSF18
Investigating the action mechanism for a natural active compound honokiol by quantitative proteomics
Imran Sajid 1,2,∗ , Khaled A. Shaaban 1 , Holm Frauendorf 1 , Shahida Hasnain 2 , Hartmut Laatsch 1
Shufang Liang 1,∗ , Yuhuan Xu 1 , Bo Ling 2 , Xia Zhao 2 , Jin Zhou 1 , Lijuan Chen 1 , Yuquan Wei 1
1
Institute of Organic and Biomolecular Chemistry, Tammanstrasse 2, Georg-August University of Gottingen 37077, Germany 2 Department of Microbiology and Molecular Genetics, University of the Punjab, Quaid-e-Azam Campus, Lahore 54590, Pakistan
1
Thiopeptide antibiotics are forming a group of cyclic peptides characterized by several common structural features, such as oxazole and thiazole units and unusual amino acids; especially dehydroamino acids are typical (Chiu et al., 1999). Thiopeptides have been used as antibacterial agents against Gram-positive bacteria and anaerobes, including pathogens resistant to antibiotics currently in use (Nagai et al., 2003), and also have potential as growth inhibitors of the human malaria parasite (Rogers et al., 1998). ValGeninthiocin (1), a new member of thiopeptide antibiotics, was isolated from the mycelium of Streptomyces sp. RSF18, along with the closely related geninthiocin (2) and the macrolide, chalcomycin. Compound 1 showed potent activity against Gram-positive bacteria and minor antifungal activity, while it was not effective against Gram-negative bacteria or microoalgae. By intensive NMR and MS studies, structure of 1 was established as a thiopeptide containing several oxazole and thiazole units and a number of unusual amino acids. We are describing here the fermentation, isolation and structure elucidation as well as the biological activity of 1.
E-mail address: [email protected] (S. Liang).
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China 2 West China Second Hospital, Sichuan University, Chengdu 610041, China
Chiu, M.L., Folche, M., Katoh, T., Puglia, A.M., Vohradsky, J., Yun, B.S., Seto, H., Thompson, C.J., 1999. Broad spectrum thiopeptide recognition specificity of the Streptomyces lividans TipAL protein and its role in regulating gene expression. J. Biol. Chem. 274, 20578–20586. Nagai, K., Kamigiri, K., Arao, N., Suzummura, K., Kawano, Y., Yamaoka, M., Zhang, H., Watanabe, M., Zuzki, K., 2003. YM-266183 and YM-266184, novel thiopeptide antibiotics produced by Bacillus cereus isolated from a Marine Sponge. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological properties. J. Antibiot. 56, 123–128. Rogers, M.J., Cundliffe, E.T., McCutchan, F., 1998. The antibiotic micrococcin is a potent inhibitor of growth and protein synthesis in the malaria parasite. Antimicrob. Agents Chemother. 42, 715–716.
The quantitative proteomics brings about a new method to investigate the action mechanism of natural active compounds from organisms. The SILAC (stable-isotope labelling by amino acids in cell culture) combined with mass spectrometry (MS) has emerged as a simple and powerful quantitative proteomic technique (Ong et al., 2002). Honokiol (HNK), an active component purified from Magnolia officinalis, a plant used in traditional Chinese, exhibits antitumor effects by inhibiting tumor growth (Bai et al., 2003), while proteins involved in antitumor activity in proteomic level are still unclear. In our study, HNK could inhibit cell proliferation and induce apoptosis of HeLa and HepG2 cells in a concentration- and time-dependent manner. We applied the SILAC–MS technique to analyze the differential proteome profiling of cells treated by HNK to investigate key proteins responsible for HNK activities. The changed proteins covered a broad variety of cellular functions including metabolism, signal transduction etc., which indicated HNK performs cytotoxicity to tumor cells through co-operating of many proteins and different pathways. Among these changed proteins, IQGAP1, -tubin, peroxiredoxin-6 and HSP70 etc. proteins were down-regulated significantly, while proteins including annexin A2 etc. were upregulated after HNK treatment. Since IQGAP1 plays important roles in cell adhesion and migration (Noritake et al., 2005), we supposed that HNK may have effects on cell migration through IQGAP1 based on our MS datasheet. Our further scratch migration assay showed that the migration inhibition of HepG2 cells can be induced by HNK, the RNA and protein expression level of IQGAP1 were respectively decreased obviously in HepG2 cells exposed to 10 g/ml HNK for 24 h. Therefore, the down-regulated expression of IQGAP1 by HNK treatment was correlated with cell migration, and HNK probably inhibits cell proliferation and migration through IQGAP1 expression changes and its interactions with other proteins.
doi:10.1016/j.jbiotec.2008.07.169
References
Keywords: Val-Geninthiocin; Thiopeptide antibiotic; Streptomyces References
Bai, X., Cerimele, F., Ushio-Fukai, M., Waqas, M., Campbell, P.M., Govindarajan, B., Der, C.J., Battle, T., Frank, D.A., Ye, K., Murad, E., Dubiel, W., Soff, G., Arbieser, J.L., 2003. Honokiol, a small molecular weight natural product, inhibits angiogenesis in vitro and tumor growth in vivo. J. Biol. Chem. 278, 35501–35507. Noritake, J., Watanabe, T., Sato, K., Wang, S., Kaibuchi, K., 2005. IQGAP1: a key regulator of adhesion and migration. J. Cell Sci. 118, 2085–2092. Ong, S.E., Blagoev, B., Kratchmarova, I., Kristensen, D.B., Steen, H., Pandey, A., Mann, M., 2002. Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics. Mol. Cell. Proteomics 1, 376–386.
doi:10.1016/j.jbiotec.2008.07.170
∗ Corresponding author at: Institute of Organic and Biomolecular Chemistry, Tammanstrasse 2, Georg-August University of Gottingen 37077, Germany.