title: Soluble fusion expression, characterization and localization of human β-defensin 6.
abstract: Human β‑defensin 6 (DEFB106) is an antimicrobial peptide expressed in the epididymis, testis and lung, which indicates that DEFB106 may be involved in innate immunity and fertility. However, as a β‑defensin, this protein has not been well characterized. Using an intein‑mediated fusion expression system, the recombinant DEFB106 was expressed and purified (yield, 3‑5 mg/l) under optimized conditions. The purified protein was characterized using mass spectrometry and circular dichroism spectroscopy. The measured molecular weight was consistent with its theoretical value and the predominant secondary structure was β‑sheet, the common structure of β‑defensin family members. The purified DEFB106 showed antimicrobial activity against not only Escherichia coli (E. coli) and Candida albicans (C. albicans) SC5314, but also Staphylococcus aureus (S. aureus) CMCC26003. Furthermore, it exhibited a high affinity for heparin and lipopolysaccharide. In addition, it was determined that native DEFB106 was located in the epididymis, bone marrow and skin. These observations may aid in the determination of the physiological and pathological functions of DEFB106.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, P.R. China.
authors.forename: Aijie
authors.initials: A
authors.lastname: Xin
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhao
authors.forename: Heguo
authors.initials: H
authors.lastname: Yu
authors.forename: Huijuan
authors.initials: H
authors.lastname: Shi
authors.forename: Haiyan
authors.initials: H
authors.lastname: Liu
authors.forename: Hua
authors.initials: H
authors.lastname: Diao
authors.forename: Yonglian
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects; D002942:Circular Dichroism; D004926:Escherichia coli / Q000187:drug effects; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013211:Staphylococcus aureus / Q000187:drug effects; D046888:Tissue Array Analysis
chemical_list: D023181:Antimicrobial Cationic Peptides; C476019:DEFB106; D010455:Peptides; D011993:Recombinant Fusion Proteins