title: Expression of a novel dual-functional protein--the antimicrobial peptide LL-37 fused with human acidic fibroblast growth factor in Escherichia coli.
abstract: Human acidic fibroblast growth factor (haFGF) stimulates repair of delayed healing which still remains a tremendously world-wide issue. However, most of the patients with delayed healings have to face another creeping problem - microbial infection, which is one of the most frequent complications that still lead to wound healing failure. LL-37/hCAP-18 is the only cathelicidin-derived antimicrobial peptide found in human with a wide range of antimicrobial activities. In the present study, a novel hybrid protein combining LL-37 with haFGF was designed. The DNA sequence encoding recombination fusion protein LL-37-haFGF was subcloned into the pET-21b vector for protein expression in Escherichia coli strain BL21 (DE3). The recombinant protein was expressed as a His-tagged protein and purified using a combination of Ni affinity and CM-Sepharose chromatography at a purity of 95.43% as detected by RP-HPLC and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Antimicrobial activity assays showed that the purified LL-37-haFGF had improved antimicrobial activities in vitro compared with LL-37. Methylthiazoletetrazolium (MTT) assay showed that the purified LL-37-haFGF also had a distinct mitogenic activity in NIH 3T3 cells. These data suggests the recombinant protein LL-37-haFGF has pharmaceutical potential for applications in wound healing.
authors.affiliation: Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China; School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, People's Republic of China; School of Life Science and Bio-pharmaceutical, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China. Electronic address: Shenjuan0412@126.com.
authors.forename: Juan
authors.initials: J
authors.lastname: Shen
authors.affiliation: Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China; School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Xue-Mei
authors.initials: XM
authors.lastname: Lu
authors.affiliation: Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China.
authors.forename: Xiao-Bao
authors.initials: XB
authors.lastname: Jin
authors.affiliation: Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Ding
authors.affiliation: Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China.
authors.forename: Xiao-Bo
authors.initials: XB
authors.lastname: Li
authors.affiliation: Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China.
authors.forename: Han-Fang
authors.initials: HF
authors.lastname: Mei
authors.affiliation: Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China.
authors.forename: Fu-Jiang
authors.initials: FJ
authors.lastname: Chu
authors.affiliation: Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, Guangdong 510006, People's Republic of China. Electronic address: zhujy@gdpu.edu.cn.
authors.forename: Jia-Yong
authors.initials: JY
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides; D001419:Bacteria / Q000187:drug effects; D054804:Cathelicidins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D016220:Fibroblast Growth Factor 1 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D006801:Humans; D051379:Mice; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology
chemical_list: D023181:Antimicrobial Cationic Peptides; D054804:Cathelicidins; D011993:Recombinant Fusion Proteins; D016220:Fibroblast Growth Factor 1