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Molecular modeling, expression and wound-repair activity in vivo of platelet-derived growth factor from Periplaneta americana
Process Biochemistry ( IF 3.7 ) Pub Date : 2021-08-12 , DOI: 10.1016/j.procbio.2021.08.009
Kaimin Lu 1 , Chuanfang Wu 1 , Jinku Bao 1
Affiliation  

Platelet-derived growth factors (PDGFs) promote the repair of the wounded tissue by stimulating macrophages to produce and secrete growth factors. Active constituents extracted from Periplaneta americana were primary for treatment of wounds, pressure sores, and burns. In this study, PDGF from Periplaneta americana (PAPDGF) was identified, and the sequence analysis, physical properties, 3D structure of PAPDGF and the interfaces of PAPDGF-PDGFRβ were predicted by bioinformatics. In addition, to explore the relationship between the key amino acid residues and activity of PAPDGF, recombinant mutant of PAPDGF (rmPAPDGF, D164H) was constructed. The sequence of PAPDGF and mutant of PAPDGF (D164H) were expressed and purified by Ni-affinity chromatography. The wound-repair activity of recombinant PAPDGF (rPAPDGF) and rmPAPDGF were tested in vivo. The results showed that rPAPDGF and rmPAPDGF were successfully obtained and exhibited a single band on SDS-PAGE at 49.87 kDa. The rPAPDGF showed potent repair activity by stimulating the expression of collagen and PDGFRβ, whereas the rmPAPDGF exhibited low repair activity in vivo, which indicated that the residue D164 in PAPDGF was the key amino acid to maintain activity. Taken together, PAPDGF may be served as a promising candidate agent for the treatment of wound in the future.



中文翻译:

美洲大蠊血小板衍生生长因子的分子建模、表达和体内伤口修复活性

血小板衍生生长因子 (PDGF) 通过刺激巨噬细胞产生和分泌生长因子来促进受伤组织的修复。从美洲大蠊中提取的活性成分主要用于治疗伤口、褥疮和烧伤。在本研究中,美洲大蠊的PDGF(PAPDGF)被鉴定,并通过生物信息学预测PAPDGF的序列分析、物理性质、3D结构和PAPDGF-PDGFRβ的界面。此外,为了探讨PAPDGF的关键氨基酸残基与活性的关系,构建了PAPDGF的重组突变体(rmPAPDGF,D164H)。PAPDGF和PAPDGF突变体(D164H)的序列通过Ni亲和层析进行表达和纯化。在体内测试了重组 PAPDGF (rPAPDGF) 和 rmPAPDGF 的伤口修复活性。结果表明,成功获得了rPAPDGF和rmPAPDGF,在SDS-PAGE上显示49.87 kDa的单条带。rPAPDGF 通过刺激胶原蛋白和 PDGFRβ 的表达显示出有效的修复活性,而 rmPAPDGF 显示出低修复活性体内,这表明PAPDGF中的残基D164是维持活性的关键氨基酸。综上所述,PAPDGF 可能成为未来治疗伤口的有希望的候选药物。

更新日期:2021-08-17
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