Study of the interaction between collagen and collagenase molecules by Dynamic Light Scattering in the presence of various modifiers
Proteins perform a lot of vital functions in living organisms: they serve as a material for building cells, tissues and organs, most hormones, hemoglobin and other compounds.
Collagenase is currently used in drugs for the treatment of diseases that result from excessive deposition of collagen protein in some organs, leading to disruption of their normal functioning [1,2]. The most commonly used bacterial collagenase derived from Clostridium histolyticum CHC (the causative agent of gas gangrene) . Since collagen makes up one third of all proteins in the human body, a violation of its synthesis can lead to diseases such as Dupuytren's syndrome, Peyronie's disease, etc. [4, 5]. In such cases, the use of an injectable form of collagenase or preparations based on it can help in the degradation of obstructive collagen.
However, there are also such diseases in which collagen fibers begin to be arbitrarily destroyed: osteochondrosis, arthrosis, visual impairment, vascular and skin conditions . Protein degradation can be slowed down by using enzyme inhibitors or protein tanning agents. Chromium plating (or tanning) of collagen has long been used not only in the leather industry, but also to stabilize and strengthen the structure of protein molecules.
The aim of this work is to investigate the interaction between collagen and collagenase molecules in the presence of various activators, inhibitors and tanning agents by dynamic light scattering in buffer solutions. It is necessary to identify and analyze the nature of the interaction as a function of time. The use of calcium, zinc, magnesium and chromium salts in this work is due to the influence on the activity of the enzyme of collagenase and on the structure of collagen fibers .
As a result of the preparatory work, it was revealed that the ions of calcium, zinc, magnesium and chromium do not significantly affect the collagen and collagenase molecules in buffer solutions separately. The obtained pH dependences of the translational diffusion coefficient Dt and hydrodynamic radius Rh for the molecules under study have a nonlinear form (close to parabolic) with minimum and maximum at the isoelectric point, respectively (pI 6.0).
Based on the results obtained, it can be concluded that chromium ions really strengthen the collagen structure, and the rate of breakdown of protein molecules is reduced by 70%. The reaction rate can be increased by adding calcium or magnesium ions to the solutions. However, the rate of biodegradation of collagen fibers that is observed in a pure solution is still not observed. The maximum rate of breakdown of protein molecules is achieved by the activation of collagenase by Ca2+ ions.
1. R. Bauer, J.J. Wilson, S.T.L. Philominathan, D. Davis, O. Matsushita, J. Sakon. Structural comparison of ColH and ColG collagen-binding domains from Clostridium histolyticum. J Bacteriol, 195 (2) (2013), pp. 318-327
2. P. Schlage, T. Kockmann, J.N. Kizhakkedathu. Monitoring matrix metalloproteinase activity at the epidermal-dermal interface by SILAC-iTRAQ-TAILS Proteomics, 15 (14) (2015), pp. 2491-2502
3. Hamzeh Alipour, Abbasali Raz, Sedigheh Zakeri, Navid Dinparast Djadid. Therapeutic applications of collagenase (metalloproteases): A review. Asian Pacific Journal of Tropical Biomedicine. Volume 6, Issue 11, November 2016, Pages 975-981
4. Nanchahal J, Midwood KS, inventors; Isis Innovation Limited, assignee. Treatment for dupuytren’s disease. United States patent US 9138458 B2. 2015 Sep 22.
5. T.C. Peak, G.C. Mitchell, F.A. Yafi, W.J. Hellstrom. Role of collagenase Clostridium histolyticum in Peyronie's disease. Biologics, 9 (2015), pp. 107-116
6. Shrutal Narendra Deshmukh, Alka M Dive,1 Rohit Moharil,1 and Prashant Munde // Enigmatic insight into collagen. J Oral Maxillofac Pathol. 2016 May-Aug; 20(2): 276–283.
7. Sergeeva I.A., Khitrina K.A., Krot A.R., Sukneva A.V., Petrova G.P. Study of the interaction and dynamics of molecules in collagen and collagenase solutions by dynamic light scattering // Izv. Sarat. un-that. New ser. Ser. Physics. 2017.Vol. 17, no. 3. S. 171–178. DOI: 10.18500 / 1817-3020-2017-17-3-171-178.
File with report