Applied Biochemistry and Biotechnology

, Volume 186, Issue 1, pp 256–270 | Cite as

Small Ubiquitin-Like Modifier Protein 3 Enhances the Solubilization of Human Bone Morphogenetic Protein 2 in E. coli

  • Muhammad Umair Hanif
  • Adnan Yaseen
  • Roquyya GulEmail author
  • Muhammad Usman Mirza
  • Muhammad Hassan Nawaz
  • Syed Shoaib Ahmed
  • Salman Aziz
  • Saima Chaudhary
  • Ayyaz Ali Khan
  • Muhammad Shoaib


Small ubiquitin-like modifier (SUMO) fusion technology is widely used in the production of heterologous proteins from prokaryotic system to aid in protein solubilization and refolding. Due to an extensive clinical application of human bone morphogenetic protein 2 (hBMP2) in bone augmentation, total RNA was isolated from human gingival tissue and mature gene was amplified through RT-PCR, cloned (pET21a), sequence analyzed, and submitted to GenBank (Accession no. KF250425). To obtain soluble expression, SUMO3 was tagged at the N-terminus of hBMP2 gene (pET21a/SUMO3-hBMP2), transferred in BL21 codon+, and ~ 40% soluble expression was obtained on induction with IPTG. The dimerized hBMP2 was confirmed with Western blot, native PAGE analysis, and purified by fast protein liquid chromatography with 0.5 M NaCl elution. The cleavage of SUMO3 tag from hBMP2 converted it to an insoluble form. Computational 3D structural analysis of the SUMO3-hBMP2 was performed and optimized by molecular dynamic simulation. Protein-protein interaction of SUMO3-hBMP2 with BMP2 receptor was carried out using HADDOCK and inferred stable interaction. The alkaline phosphatase assay of SUMO3-hBMP2 on C2C12 cells showed maximum 200-ng/ml dose-dependent activity. We conclude that SUMO3-tagged hBMP2 is more suited for generation of soluble form of the protein and addition of SUMO3 tag does not affect the functional activity of hBMP2.


hBMP2 SUMO3 Rosetta gami B(DE3) BL21 codon+ Fast protein liquid chromatography Alkaline phosphatase 



The authors greatly acknowledge Dr. Muhammad Waheed Akhtar, Director School of Biological Sciences and his team from University of the Punjab, Lahore for their enormous support in the expression and purification experiments.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12010_2018_2736_MOESM1_ESM.docx (536 kb)
ESM 1 (DOCX 536 kb)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Muhammad Umair Hanif
    • 1
    • 2
  • Adnan Yaseen
    • 1
  • Roquyya Gul
    • 1
    • 2
    • 3
    Email author
  • Muhammad Usman Mirza
    • 4
  • Muhammad Hassan Nawaz
    • 1
  • Syed Shoaib Ahmed
    • 1
    • 2
  • Salman Aziz
    • 3
    • 5
  • Saima Chaudhary
    • 3
    • 6
  • Ayyaz Ali Khan
    • 3
    • 7
  • Muhammad Shoaib
    • 1
    • 2
    • 3
  1. 1.Institute of Molecular Biology and Biotechnology/Centre for Research in Molecular MedicineThe University of LahoreLahorePakistan
  2. 2.Centre for Research in Molecular MedicineThe University of LahoreLahorePakistan
  3. 3.Division of Molecular MedicineInstitute of Advanced Dental Sciences and ResearchLahorePakistan
  4. 4.Department of Pharmaceutical Sciences, Rega Institute for Medical ResearchUniversity of LeuvenLeuvenBelgium
  5. 5.Faculty of Medical and Allied Health SciencesSuperior UniversityLahorePakistan
  6. 6.Department of Oral PathologyUniversity of Health SciencesLahorePakistan
  7. 7.Department of Oral Health, Shaikh Zayed Medical ComplexFederal Postgraduate Medical InstituteLahorePakistan

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