PERIPLASMIC EXPRESSION OF A NOVEL HUMAN BONE MORPHOGENETIC PROTEIN-7 MUTANT IN ESCHERICHIA COLI

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NEMATOLLAHI LEILA; KHALAJ VAHID; BABAZADEH SEYEDEH MALIHEH; RAHIMPOUR AZAM; JAHANDAR HODA; DAVAMI FATEMEH; MAHBOUDI FEREIDOUN

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Journal: Avicenna Journal of Medical Biotechnology Year:2012 | Volume:4 | Issue:4 (15) Page(s): 178-185

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Title

PERIPLASMIC EXPRESSION OF A NOVEL HUMAN BONE MORPHOGENETIC PROTEIN-7 MUTANT IN ESCHERICHIA COLI

Author(s)

Keywords

BONE MORPHOGENETIC PROTEIN-7 (BMP-7)

ESCHERICHIA COLI

PERIPLASMIC EXPRESSION

Abstract

Background: Bone Morphogenetic Proteins (BMPs) belong to the transforming growth factor-b (TGF-b) superfamily, and play an important role in bone metabolism. Recombinant forms of BMP-2 and BMP-7 are the only BMPs used clinically. In this study the mature part of human bone morphogenetic protein-7 (BMP-7) was engineered through substitution of the BMP-7 N terminal sequence by heparin-binding site of BMP-2. This targeted substitution was made to enhance the binding affinity of the novel protein to the extracellular matrix components such as heparin and heparan sulfate proteoglycans (HSPGs).Methods: The engineered protein was expressed in ESCHERICHIA COLI (E. coli). The Pel B signal sequence was used to translocate soluble proteins into the periplasmic space of E. coli. The protein was purified from periplasmic extract using Ni-NTA chromatography and the SDS-PAGE and western blot analysis confirmed the successful expression of the novel protein.Results: The novel hBMP-7 mutant was produced as approximately 16 kDa monomer. It was found that the heparin binding of this protein was approximately 50% more than that of the wild-type at a protein concentration of 500ng/ml.Conclusion: The findings showed that the PERIPLASMIC EXPRESSION may be suitable to produce complex proteins like BMPs.

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APA: Copy

NEMATOLLAHI, LEILA, KHALAJ, VAHID, BABAZADEH, SEYEDEH MALIHEH, RAHIMPOUR, AZAM, JAHANDAR, HODA, DAVAMI, FATEMEH, & MAHBOUDI, FEREIDOUN. (2012). PERIPLASMIC EXPRESSION OF A NOVEL HUMAN BONE MORPHOGENETIC PROTEIN-7 MUTANT IN ESCHERICHIA COLI. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), 4(4 (15)), 178-185. SID. https://sid.ir/paper/313792/en

Vancouver: Copy

NEMATOLLAHI LEILA, KHALAJ VAHID, BABAZADEH SEYEDEH MALIHEH, RAHIMPOUR AZAM, JAHANDAR HODA, DAVAMI FATEMEH, MAHBOUDI FEREIDOUN. PERIPLASMIC EXPRESSION OF A NOVEL HUMAN BONE MORPHOGENETIC PROTEIN-7 MUTANT IN ESCHERICHIA COLI. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)[Internet]. 2012;4(4 (15)):178-185. Available from: https://sid.ir/paper/313792/en

IEEE: Copy

LEILA NEMATOLLAHI, VAHID KHALAJ, SEYEDEH MALIHEH BABAZADEH, AZAM RAHIMPOUR, HODA JAHANDAR, FATEMEH DAVAMI, and FEREIDOUN MAHBOUDI, “PERIPLASMIC EXPRESSION OF A NOVEL HUMAN BONE MORPHOGENETIC PROTEIN-7 MUTANT IN ESCHERICHIA COLI,” AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), vol. 4, no. 4 (15), pp. 178–185, 2012, [Online]. Available: https://sid.ir/paper/313792/en

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