Identification and Characterisation of a Novel Protein FIP-sch3 from Stachybotrys chartarum

abstract: ...In this study, a novel FIP named FIP-sch3 has been identified and characterised. FIP-sch3 was identified in the ascomycete Stachybotrys chartarum , making it the second FIP to be identified outside the order of Basidiomycota. Recombinant FIP-sch3 (rFIP-shc3) was produced in Escherichia coli and purified using GST-affinity magnetic beads. The bioactive characteristics of FIP-sch3 were compared to those of well-known FIPs LZ-8 from Ganoderma lucidum and FIP-fve from Flammulina velutipes , which were produced and purified using the same method. The purified rFIP-sch3 exhibited a broad spectrum of anti-tumour activity in several types of tumour cells but had no cytotoxicity in normal human embryonic kidney 293 cells. Assays that were implemented to study these properties indicated that rFIP-sch3 significantly suppressed cell proliferation, induced apoptosis and inhibited cell migration in human lung adenocarcinoma A549 cells. The anti-tumour effects of rFIP-sch3 in A549 cells were comparable to those of rLZ-8, but they were significantly greater than those of rFIP-fve. Molecular assays that were built on real-time PCR further revealed potential mechanisms related to apoptosis and migration and that underlie phenotypic effects. These results indicate that FIP-shc3 has a unique anti-tumour bioactive profile, as do other FIPs, which provide a foundation for further studies on anti-tumour mechanisms. Importantly, this study also had convenient access to FIP-sch3 with potential human therapeutic applications.

...PLoS One PLoS ONE plos plosone PLoS ONE 1932-6203 Public Library of Science San Francisco, CA USA 27997578 5173029 10.1371/journal.pone.0168436 PONE-D-16-32084 Research Article Biology and Life Sciences Cell Biology Cell Processes Cell Death Apoptosis Research and analysis methods Biological cultures Cell lines HEK 293 cells Medicine and Health Sciences Oncology Cancer Treatment Research and Analysis Methods Database and Informatics Methods Database Searching Sequence Similarity Searching Biology and Life Sciences Genetics Gene Expression Biology and Life Sciences Toxicology Cytotoxicity Medicine and Health Sciences Pathology and Laboratory Medicine Toxicology Cytotoxicity Research and Analysis Methods Database and Informatics Methods Bioinformatics Sequence Analysis Sequence Alignment Biology and Life Sciences Cell Biology Cell Motility Cell Migration Cancer Cell Migration Biology and Life Sciences Developmental Biology Cell Migration Cancer Cell Migration Identification and Characterisation of a Novel Protein FIP-sch3 from Stachybotrys chartarum A Novel Protein FIP-sch3 from Stachybotrys chartarum Li Shuying 1 Zhao Leiming 1 Xu Wenyi 1 Jiang Zhonghao 1 Kang Jun 2 Wang Fengzhong 1 * Xin Fengjiao 1 * 1 Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing, China 2 School of Life Sciences, Tianjin University, Tianjin, China Lebedeva Irina V Editor Columbia University, UNITED STATES Competing Interests: The authors have declared that no competing interests exist. Conceptualization: SYL. Data curation: SYL. Methodology: WYX ZHJ. Project administration: SYL FJX. Resources: LMZ JK. Supervision: FZW. Validation: SYL WYX ZHJ. Writing original draft: SYL. Writing review & editing: SYL. * E-mail: wangfengzhong@sina.com (FW); 2002hongzhi30@163.com (FX) 20 12 2016 2016 11 12 e0168436 10 8 2016 1 12 2016 2016 Li et al 2016 Li et al This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. In this study, a novel FIP named FIP-sch3 has been identified and characterised. FIP-sch3 was identified in the ascomycete Stachybotrys chartarum , making it the second FIP to be identified outside the order of Basidiomycota. Recombinant FIP-sch3 (rFIP-shc3) was produced in Escherichia coli and purified using GST-affinity magnetic beads. The bioactive characteristics of FIP-sch3 were compared to those of well-known FIPs LZ-8 from Ganoderma lucidum and FIP-fve from Flammulina velutipes , which were produced and purified using the same method. The purified rFIP-sch3 exhibited a broad spectrum of anti-tumour activity in several types of tumour cells but had no cytotoxicity in normal human embryonic kidney 293 cells. Assays that were implemented to study these properties indicated that rFIP-sch3 significantly suppressed cell proliferation, induced apoptosis and inhibited cell migration in human lung adenocarcinoma A549 cells. The anti-tumour effects of rFIP-sch3 in A549 cells were comparable to those of rLZ-8, but they were significantly greater than those of rFIP-fve. Molecular assays that were built on real-time PCR further revealed potential mechanisms related to apoptosis and migration and that underlie phenotypic effects. These results indicate that FIP-shc3 has a unique anti-tumour bioactive profile, as do other FIPs, which provide a foundation for further studies on anti

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Gene: SHC3 - SHC (Src homology 2 domain containing) transforming protein 3 - (NCBI)

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