Volume 20, Issue 5 (Iranian South Medical Journal 2017)
Iran South Med J 2017, 20(5): 481-491 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Hosseini S, Zolgharnine H, Bargahi A, Khaledi H, Salamat N, Archangi B. Purification of Bysal Mussel (Modiolus sp. PG) Adhesive Protein fp-2 from Nortern Seashore of Persian Gulf. Iran South Med J 2017; 20 (5) :481-491
URL: http://ismj.bpums.ac.ir/article-1-900-en.html
URL: http://ismj.bpums.ac.ir/article-1-900-en.html
Saba Hosseini1 
, Hossean Zolgharnine * 2, Afshar Bargahi3 , Hoda Khaledi1 , Negin Salamat1 , Bita Archangi1
, Hossean Zolgharnine * 2, Afshar Bargahi3 , Hoda Khaledi1 , Negin Salamat1 , Bita Archangi1
1- Department of Marine Biology, School of Marine Science and Ocean, Khorramshahr University of Marine Science and Technology, Khorramshahr, Iran
2- Department of Marine Biology, School of Marine Science and Ocean, Khorramshahr University of Marine Science and Technology, Khorramshahr, Iran ,Zolgharnine@yahoo.com
3- The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
2- Department of Marine Biology, School of Marine Science and Ocean, Khorramshahr University of Marine Science and Technology, Khorramshahr, Iran ,
3- The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
Abstract: (4568 Views)
Background: Mussel bivalves produce and secrete adhesive proteins which facilitate the adherence to surfaces in the aquatic environments. Specific properties of these proteins such as adhesion, resistance and biocompatibility have introduced their application in biomedical engineering, dental, and production of biological adhesives.
Materials and Methods: The species of dwarves from the Persian Gulf were sampled based on the morphologic indexes and COI gene sequence. To identify the polyphenol glands, which produce the adhesive proteins, tissue sections were prepared from the mussel’s foot. Then, the mussel fp-2 protein was purified by the ion-exchange chromatography using salt and pH gradients and, the gel filtration chromatography. To confirm the presence of proteins in each fraction the Arnow test and NBT staining were used. For detection of the protein homogeneity, in each purification stage, the SDS-PAGE electrophoresis was used. The NBT positive chromatogram peaks, after concentration and electrophoresis by Acid-Urea method, were blotted on nitrocellulose and detected by NBT test.
Results: The specie morphology and taxonomy were recorded and matched to the FAO criteria and, "The Mollusks of Persian Gulf", the Persian reference book. It belonged to the Mytilidae family and Modiolus genus. The DNA and protein sequences were recorded in NCBI data bank as AB972412.1 and BAP16437.1 respectively. Also, the molecular weight of the purified fp-2 protein was about 45 KD.
Conclusion: This research was led to the morphologic and genetic identification of Modiolus sp. PG from the Persian Gulf along with the purification of the fp-2 adhesive protein.
Materials and Methods: The species of dwarves from the Persian Gulf were sampled based on the morphologic indexes and COI gene sequence. To identify the polyphenol glands, which produce the adhesive proteins, tissue sections were prepared from the mussel’s foot. Then, the mussel fp-2 protein was purified by the ion-exchange chromatography using salt and pH gradients and, the gel filtration chromatography. To confirm the presence of proteins in each fraction the Arnow test and NBT staining were used. For detection of the protein homogeneity, in each purification stage, the SDS-PAGE electrophoresis was used. The NBT positive chromatogram peaks, after concentration and electrophoresis by Acid-Urea method, were blotted on nitrocellulose and detected by NBT test.
Results: The specie morphology and taxonomy were recorded and matched to the FAO criteria and, "The Mollusks of Persian Gulf", the Persian reference book. It belonged to the Mytilidae family and Modiolus genus. The DNA and protein sequences were recorded in NCBI data bank as AB972412.1 and BAP16437.1 respectively. Also, the molecular weight of the purified fp-2 protein was about 45 KD.
Conclusion: This research was led to the morphologic and genetic identification of Modiolus sp. PG from the Persian Gulf along with the purification of the fp-2 adhesive protein.
Type of Study: Original |
Subject:
Disorders of Systemic, Metabolic or Environmental Origin
Received: 2016/12/29 | Accepted: 2017/04/19 | Published: 2017/11/13
Received: 2016/12/29 | Accepted: 2017/04/19 | Published: 2017/11/13
References
1. Tsang PH, Li G, Brun YV, et al. Adhesion of single bacterial cell in micronewton range. Proc Natl Acad Sci USA 2006; 103(15): 5764-8. [DOI:10.1073/pnas.0601705103]
2. Vollrath F. Strength and structure of spiders' silks. J Biotechnol 2000; 74(2): 67-83. [DOI:10.1016/S1389-0352(00)00006-4]
3. Stewart RJ, Weaver JC, Morse DE, et al. The tube cement of Pharamatopoma californica: a solid foam. J Exp Biol 2004; 207(Pt 26): 4727-34. [DOI:10.1242/jeb.01330]
4. DeMoor S, Waite JH, Jangoux M, et al. Characterization of the adhesive from cuvierian tubules of the sea cucumber holothuria forskali (echinodermata, holothuroidea). Mar Biotechnol (NY) 2003; 5(1): 45-57. [DOI:10.1007/s10126-002-0049-2]
5. Kamino K, Inoue K, Maruyama T, et al. Barnacles cement proteins. Importance of disulfide bonds in their insolubility. J Biol Chem 2000; 275(35): 27360-5. [DOI:10.1074/jbc.M910363199]
6. Silva SS, Mano JF, Reis RL. Potential applications of natural origin polymer-based systems in soft tissue regeneration. Crit Rev Biotechnol 2010; 30(3):200-21. [DOI:10.3109/07388551.2010.505561]
7. Heim M, Römer L, Scheibel T. Hierarchical structures made of proteins. The complex architecture of spider webs and their constituent silk proteins. Chem Soc Rev 2010; 39(1): 156-64. [DOI:10.1039/B813273A]
8. Lee BP, Messersmith PB, Israelachvili JN, et al. Mussel-inspired adhesives and coatings. Annu Rev Mater Res 2011; 41: 99-132. [DOI:10.1146/annurev-matsci-062910-100429]
9. Strausberg RL, Link RP. Protein-based medical adhesives. Trends Biotechnol 1990; 8(2): 53-7. [DOI:10.1016/0167-7799(90)90134-J]
10. Cranford SW, Tarakanova A, Pugno NM, et al. Nonlinear material behaviour of spider silk yields robust webs. Nature 2012; 482(7383): 72-6. [DOI:10.1038/nature10739]
11. Z, Cranford S, Ackbarow T, et al. Robustness-strength performance of hierarchical alpha-helical protein filaments. Int J Appl Mech 2009; 1(1): 85-112. [DOI:10.1142/S1758825109000058]
12. Sen D, Buehler MJ. Structural hierarchies define toughness and defect-tolerance despite simple and mechanically inferior brittle building blocks. Sci Rep 2011; 1: 35. [DOI:10.1038/srep00035]
13. Waite JH. The DOPA ephemera: a recurrent motif in invertebrates. Biol Bull 1992; 183(1): 178-84. [DOI:10.2307/1542421]
14. Qin Z, Buehler MJ. Impact tolerance in mussel thread networks by heterogeneous material distribution. Nat Commun 2013; 4: 2187. [DOI:10.1038/ncomms3187]
15. Deming TJ. Mussel byssus and biomolecular materials. Curr Opin Chem Biol 1999; 3(1): 100-5. [DOI:10.1016/S1367-5931(99)80018-0]
16. DS, Gim Y, Yoo HJ, et al. Practical recombinant hybrid mussel bioadhesive fp-151. Biomaterials 2007; 28(24): 3560-8. [DOI:10.1016/j.biomaterials.2007.04.039]
17. Lim S, Choi YS, Kang DG, et al. The adhesive properties of coacervated recombinant hybrid mussel adhesive proteins. Biomaterials 2010; 31(13): 3715-22. [DOI:10.1016/j.biomaterials.2010.01.063]
18. Murphy JL, Vollenweider L, Xu F, et al. Adhesive performance of biomimetic adhesive-coated biologic scaffolds. Biomacromolecules 2010; 11(11): 2976-84. [DOI:10.1021/bm1007794]
19. Waite JH. Evidence for a repeating 3, 4-dihydroxyphenylalanin-contaning and hydroxyproline-containing decapeptide in the adhesive protein of the mussel Mytilus edulis. J Biol Chem 1983; 258(5): 2911-5.
20. Silverman HG, Roberto FF. Understanding marine mussel adhesion. Mar Biotechnol (NY) 2007; 9(6): 661-81. [DOI:10.1007/s10126-007-9053-x]
21. Lawlor EP. Discover Nature at the Seashore: Things to know and things to do. Mechanicsburg, Pennsylvania: Stackpole Book; 1992.
22. Ragnarsson SA, Burgos JM. Separating the effects of a habitat modifier, Modiolus modiolus and substrate properties on the associated megafauna. Journal of Sea Research 2012; 72: 55–63. [DOI:10.1016/j.seares.2012.05.011]
23. Danner EW, Kan Y, Hammer MU, et al. Adhesion of mussel foot protein Mefp-5 to mica: an underwater superglue. Biochemistry 2012; 51(33): 6511-8. [DOI:10.1021/bi3002538]
24. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-54. [DOI:10.1016/0003-2697(76)90527-3]
25. Ninan L, Monahan J, Storshine RL, et al. Adhesive strength of marine mussel extracts of porcine skin. Biomaterial 2003; 24(22): 4091-9. [DOI:10.1016/S0142-9612(03)00257-6]
26. Arnow LE. Colorimetric determination of the components of 3, 4 dihydroxyphenylalanine tyrosine mixtures. J Biol Chem 1937; 118(2): 531-7.
27. Paz MA, Flückiger R, Boak A, et al. Specific detection of quinoproteins by redox-cycling staining. J Biol Chem 1991; 266(2): 689-92.
28. Zhao H, Waite JH. Linking adhesive and structural proteins in the attachment plaque of Mytilus californianus. J Biol Chem 2006; 281(36): 26150-8. [DOI:10.1074/jbc.M604357200]
29. Zhao H, Waite JH. Proteins in load-bearing junctions: The histidine-rich metal-binding protein of mussel byssal. Biochemistry 2006; 45(47):14223-31. [DOI:10.1021/bi061677n]
30. Walker JM. p The Protein Protocols Handbook. Humana Press, a Part of Springer Science. 1985.
31. Brazee, SL, Carrington E, Interspecific comparison of the mechanical properties of mussel byssus. Biological Bulletin. 2006; 211, 263–274 [DOI:10.2307/4134548]
32. Hwang DS, Sim SB, Cha HJ. Cell adhesion biomaterial based on mussel adhesive protein fused with RGD peptide. Biomaterials 2007; 28(28):4039-46. [DOI:10.1016/j.biomaterials.2007.05.028]
Send email to the article author
| Rights and Permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
