Skip to main content

Advertisement

SpringerLink
Log in

Urea-Induced Dissociation of Nerve Growth Factor from Venom of Chinese Cobra by SEC

  • Original
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

The urea-induced dissociation of nerve growth factor from venom of Chinese cobra (cNGF) was studied by intrinsic fluorescence emission spectra, SEC, urea-gradient polyacrylamide gel electrophoresis, assays of biological activity and thermodynamic parameters. The results showed that when urea concentration was lower than or equal to 4.0 mol L−1 or higher than or equal to 8.0 mol L−1, cNGF existed only in native homodimer form or monomer form, respectively; whereas when urea concentration was higher than 4.0 mol L−1 and lower than 8.0 mol L−1, they existed simultaneously in the native homodimer and monomer forms and the former decreased, while the latter increased with the increase in urea concentration. Based on the association–dissociation equilibrium between cNGF and urea molecules, an equation, which includes two characteristic dissociation parameters K and ∆m, was presented to describe the urea-induced dissociation process of cNGF. As the reaction temperature increased from 15 to 35 °C, positive enthalpy and entropy changes were observed, and the parameter K increased from 2.72 × 10−13 to 5.18 × 10−12 (L mol−1), while the parameters ∆m and ∆G, respectively, decreased from 10.18 to 8.42 and from −10.27 to −18.67 (kJ mol−1), which means that the urea-induced dissociation of cNGF was spontaneous and entropy-driven and the higher temperature was favorable for the dissociation process. Using the procedures and equations mentioned in the paper, the urea-induced dissociation of cNGF is first comprehensively described. Furthermore, this work presents a useful method for people to study the dissociation of dimer or multimer proteins induced by denaturants, inducers, pH, etc.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. McCleary RJ, Kini RM (2013) Non-enzymatic proteins from snake venoms: a gold mine of pharmacological tools and drug leads. Toxicon 62:56–74

    Article  CAS  Google Scholar 

  2. Barbacid M (1995) Neurotrophic factors and their receptors. Curr Opin Cell Biol 7:148–155

    Article  CAS  Google Scholar 

  3. Levi Montalcini R (1987) The growth factor 35 years later. Science 237:1154–1162

    Article  CAS  Google Scholar 

  4. Fujita K, Lazarovici P, Guroff G (1989) Regulation of the differentiation of PC12 pheochromocytoma cells. Environ Health Perspect 80:127–142

    Article  CAS  Google Scholar 

  5. Lecht S, Arien-Zakay H, Wagenstein Y, Inoue S, Marcinkiewicz C, Lelkes PI, Lazarovici P (2010) Transient signaling of Erk1/2, Akt and PLCγ induced by nerve growth factor in brain capillary endothelial cells. Vasc Pharmacol 53:107–114

    Article  CAS  Google Scholar 

  6. Tuszynski MH, Conner J, Bleshh A, Smith D, Merrill DA, Lee Vahlsing H (2002) New strategies in neural repair. Progr Brain Res 138:401–409

    Article  CAS  Google Scholar 

  7. Capsoni S, Giannotta S, Cattaneo A (2002) Beta-amyloid plaques in a model for sporadic Alzheimer’s disease based on transgenic anti-nerve growth factor antibodies. Mol Cell Neurosci 21:15–28

    Article  CAS  Google Scholar 

  8. Hellweg R, Hartung HD (1990) Endogenous levels of nerve growth factor (NGF) are altered in experimental diabetes mellitus: a possible role for NGF in the pathogenesis of diabetic neuropathy. J Neurosci Res 26:258–267

    Article  CAS  Google Scholar 

  9. Levi-Montalcini R, Meyer H, Hamburger V (1954) In vitro experiments on the effects of mouse sarcomas 180 and 37 on the spinal and sympathetic ganglia of the chick embryo. Cancer Res 14:49–57

    CAS  Google Scholar 

  10. Kostiza T, Meier J (1996) Nerve growth factor from snake venom: chemical properties, mode of action and biological significance. Toxicon 34:787–806

    Article  CAS  Google Scholar 

  11. Siigur J, Arumae U, Neuman T, Siigur E, Saarma M (1987) Monoclonal antibody immunoaffinity chromatography of the nerve growth factor from snake venoms. Comp Biochem Physiol B 87:329–334

    CAS  Google Scholar 

  12. Thoenen H, Barde YA (1980) Physiology of nerve growth factor. Physiol Rev 60:1284–1335

    CAS  Google Scholar 

  13. Xu T, Wang W, Huang Y, MEng Q, Li D, Lu Q, Xiong Y (1999) A nerve growth factor from the venom of Chinese cobra (Naja naja atra) and its effects on male reproductive system in rats. Comp Biochem Phys C 124:149–156

    Article  CAS  Google Scholar 

  14. Wei J, Mo Y, Qiao L, Wei X, Chen H, Xie H, Fu Y, Wang W, Xiong Y, He S (2006) Potent histamine-releasing activity of atrahagin, a novel snake venom metalloproteinase. Int J Biochem Cell B 38:510–520

    Article  CAS  Google Scholar 

  15. Zhang Y, Zhong L, Zhou B, Chen J, Li C (2013) Interaction of characteristic structural elements of persimmon tannin with Chinese cobra PLA2. Toxicon 74:34–43

    Article  CAS  Google Scholar 

  16. Tong Q, Wang F, Zhou H, Sun H, Song H, Shu Y, Gong Y, Zhang W, Cai T, Yang F, Tang J, Jiang T (2012) Structural and functional insights into lipid-bound nerve growth factors. FASEB J 26:3811–3821

    Article  CAS  Google Scholar 

  17. Kaplan DR, Stephens RM (1994) Neurotrophin signal transduction by the trk receptor. J Neurobiol 25:1404–1417

    Article  CAS  Google Scholar 

  18. Jiang H, Movsesyan V, Fink DW, Fasler M, Whalin M, Katagiri Y, Monshipouri M, Dickens G, Lekles PI, Guroff G, Lazarovici P (1997) Expression of human p140trk receptors in p140trk-deficient, PC12/endothelial cells results in nerve growth factor-induced signal transduction and DNA synthesis. J Cell Biochem 66:229–244

    Article  CAS  Google Scholar 

  19. Katzir I, Shani J, Goshen G, Sela J, Ninary E, Dogonovski AM, Shabashov D, Inoue S, Ikeda K, Hayashi K, Gorinstein S, Deutsch J, Lazarovici P (2003) Characterization of nerve growth factors (NGFs) from snake venoms by use of a novel, quantitative bioassay utilizing pheochromocytoma (PC12) cells overexpressing human trkA receptors. Toxicon 42:481–490

    Article  CAS  Google Scholar 

  20. Bian L, Wu P, Yang X (2004) Two-step chromatographic method for separation and purification of nerve growth factor from venom of Chinese Cobra. J Chromatogr B 805:119–125

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was financially supported by National Natural Science Foundation of China (No. 21075097) and Key Program for Science and Technology Innovative Research Team of Shaanxi Province (No. 2013KCT-24).

Author information

Authors and Affiliations

  1. College of Life Science, Northwest University, 229 Taibai Road, Xi’an, 710069, China

    Liujiao Bian & Xu Ji

  2. Emergency Department, Shaan’xi Provincial People’s Hospital, 256 Youyi Road, Xi’an, 710068, China

    Wei Hu

Authors
  1. Liujiao Bian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xu Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wei Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Liujiao Bian.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bian, L., Ji, X. & Hu, W. Urea-Induced Dissociation of Nerve Growth Factor from Venom of Chinese Cobra by SEC. Chromatographia 77, 793–802 (2014). https://doi.org/10.1007/s10337-014-2661-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10337-014-2661-5

Keywords

Search

Navigation