A TAXONOMIC STUDY ON ORTHRIAS INSIGNIS EUPHRATICUS (Banarescu and Nalbant, 1964) and CYPRINION MACROSTOMUS (Heckel, 1843) by SARCOPLASMIC PROTEIN ELECTROPHORESIS

Abstract

In this study, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) was applied to the sarcoplasmic proteins of Orthrias insignis euphyraticus (BALITORIDAE) and Cyprinion macrostomus (CYPRINIDAE) fish taken from Karakaya Dam Lake. The electrophoregram showed that there were differences between the two species in both the number of bands and the molecular weight (MW) of the sarcoplasmic proteins. While Cyprinion macrostomus had 18 protein bands, there were 20 protein bands in Orthrias insignis euphyraticus. On the other hand, the bands obtained from Orthrias insignis euphyraticus and numbered as 9th protein band with molecular weight of 29.5 kD and 12th band with MW of 27.2 kD were not found in Cyprinion macrostomus. 9th protein band with MW of 27.5 kD was only found in Cyprinion macrostomus. We concluded that comparisons of sarcoplasmic proteins might have practical importance in taxonomic classification.

Keywords

• Key Words: Fish
• Sarcoplasmic Proteins
• SDS-PAGE
• Taxonomy

SARKOPLAZMİK PROTEİN ELEKTROFOREZİ İLE ORTHRIAS INSIGNIS EUPHRATICUS (Banarescu and Nalbant, 1964) ve CYPRINION MACROSTOMUS (Heckel, 1843) TAKSONOMİSİNİN İNCELENMESİ

Abstract

Bu çalışmada, SDS-PAGE (sodyum dodesil sülfat-poliakrilamid jel elektroforezi)’ne, Karakaya Baraj Gölü’nden yakalanan Orthrias insignis euphyraticus (BALITORIDAE) ve Cyprinion macrostomus (CYPRINIDAE) balıklarının sarkoplazmik proteinleri uygulandı. Elektroforegramda, sarkoplazmik proteinlerin band sayısı ve molekül ağırlıklarının iki tür arasında farklı olduğu görüldü. Cyprinion macrostomus 18 protein bandına sahipken Orthrias insignis euphyraticus’da 20 protein bandı vardı. Diğer taraftan, Orthrias insignis euphyraticus’dan elde edilen 29.5 kD moleküler ağırlığına sahip 9. protein bandı ve 27.2 kD moleküler ağırlığına sahip 12. protein bandı Cyprinion macrostomus’ta bulunmadı. 27.5 kD moleküler ağırlığına sahip 9. protein bandı yalnızca Cyprinion macrostomus’ta bulundu. Sarkoplazmik proteinlerin karşılaştırılmasının taksonomik sınıflandırmada pratik önemi olabileceğine karar verildi

Keywords

• Balık
• Sarkoplazmik proteinler
• SDS-PAGE
• Taksonomi

References

1. Focant B., Jacob M.F., and Huriaux, F., “Electrophoretic comparison of the proteins of some perch (Perca fluviatilis L.) head muscles”, J. Muscle Res. Cell Motil., 2 (3): 295-305 (1981).
2. Khoo G., Loh E.Y.F., Lim T.M., and Phang V.P.E., “Genetic variation in different varieties of siamese fighting fish using isoelectric focusing of sarcoplasmic proteins”, Aquacult. Int., 5: 537-549 (1997).
3. Pinerio C., Sotelo C.G., Medina I., Gallerdo I.M., and Martin R.P., “Reversed-phase HPLC as a method for the identification of gadoid fish species”, Z. Lebensm. Unters. Forsch. A, 204: 411-416 (1997).
4. Knuutinen J., and Harjula, P., “Identification of fish by reversed-phase high performance liquid chromatography with photodiode-array detection”, J. Chromatogr., 705B: 11-21 (1998).
5. Miyazaki J.I., Hirabayashi T., Hosoya K., and Iwami T.A., “Study of the systematics of cyprinid fishes by two dimensional gel electrophoresis”, Environ. Biol. Fish., 52: 173-179 (1998).
6. Colombo M.M., Colombo F., Biondi P.A., “Malandra R. and Renon P., “Substitution of fish species detected by thin-layer isoelectric focusing and a computer-assisted method for the evalution of gels”, Chromatogr., 880: 303-309 (2000).
7. Yılmaz M., Türköz Y., Erdemli A.Ü., Kalkan E. ve Çiğremiş Y., “Karakaya baraj gölü bazı balıklarının kan serum proteinlerinin elektroforetik modelleri üzerine taksonomik bir çalışma”, S.Ü.Vet. Bil. Derg., 16(1): 89-92 (2000).
8. Pinerio C., Barros-Velazques J., Perez-Martin R.I., Martinez I., Jacobsen T., Rehbein H., and et al. “Development of a sodium dodecyl sulfate-polyacrylamide gel electrophoresis reference method for the analysis and identification of fishy species in raw and heat-processed samples: a collaborative study”, Electrophoresis, 20: 1425-1432 (1999).

9. Pineiro C., Vazquez J., Marina A.I., Barros-Velazquez J., and Gallardo J.M., “Characterization and partial sequencing of species-specific sarcoplasmic polypeptides from commercial hake species by mass spectrometry following two-dimensional electrophoresis”, Electrophoresis, 22 (8): 1545-1552 (2001).
10. Goll D.E., Muscle proteins. Chap. 6. In “food proteins, Whitaker J.R., and Tannenbaum S.R., (Ed.) Avi Publishing Co., Wespert, CT., p.121 (1977).
11. An H., Marshall M.R., Otwell W.S., and Wei, C.I., “Electrophoretic identification of raw and cooked shrimp using various protein extraction systems”, J. Food Sci., 53 (2): 313-318 (1988).
12. Lowry O.H., Rosebrough N.J., Farr A.L., and Randall, R.J., “Protein measurement with the folin phenol reagent”, J. Biol. Chem., 193: 265-271 (1951).
13. Laemmli UK., “Cleavage of structural proteins during the assembly of the head of bacteriophage T4”, Nature, 227: 680-685 (1970).
14. O’Farrell P.H., “High resolution two-dimensional electrophoresis of proteins”, J. Biol. Chem., 250: 4007-4021 (1975).
15. Weber K., Pringle J.R., and Osborn M., “Measurement of molecular weights by electrophoresis on SDS-acrylamide gel”, Meth. Enzymol., 26: 3 (1972)”
16. ImageJ program, version 1.30 (http://rsb.info.nih.gov/ij/).
17. Lundstrom R.C. “Fish species identification by isoelectric focusing: sarcoplasmic protein polymorphism in monkfish (Lophius americanus)”, J. Assoc. Off Anal. Chem., 64 (1): 32-37 (1981).
18. Macki I.M., Pryde S.E., Sotelo C.G., Medina I., Martin R.P., Quinterio J., and Mendez M.R., “Challenge in the identification of species of canned fish”, Trends Food Sci. Tech., 10: 9-14 (1999).
19. Valenzuela M.A., Gamarra N., Gomez L., Kettlun A.M., Sardon M., Perez L.M., Vinagre J., and Guzman N.A. “A comparative study of fish species identification by gel isoelectrofocusing two- dimensional gel electrophoresis, and capillary zone electrophoresis”, J. Capillary Electrophor., 6 (3-4): 85-91 (1999).
20. Laird W.M., Mackie I.M., and Ritchie A.H., “Differentiation of species of fish by isoelectric focusing on agarose and polyacrylamide gels-a comparison”, J. Assoc. Publ. Analysis, 20: 125-135 (1982).
21. Mukhopadhyay S., Basu S., Data C., and Bose A., “Electrophoretic study on serum proteins fresh water teleosts”, Nucleus, 30 (3): 101-103 (1987).
22. Khan A.R., and Gadru M., “Electrophoretic patterns of blood serum pattern of some fish of Kashmir”, Trop. Freshwater Biol., 11: 62-70 (1988).
23. LeBlanc E.L., and LeBlanc R.J., “Capillary zone electrophoresis of fish muscle sarcoplasmic proteins”, J. Food Sci., 59 (6): 1267-1270 (1994).
24. Rehbein H., Kündiger R., Yman I.M., Ferm M., Etienne M., Jerome M., and et al., “Species identification of cooked fish by urea isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis: a collaborative study”, Food Chem., 67: 333-339 (1999).
25. LeBlanc E.L., and LeBlanc R.J., “Separation of Cod (Gadus morhua) fillet proteins by electrophoresis and HPLC after various frozen storage treatments”, J. Food Sci., 54(4): 827-834 (1989).
26. Ragnarsson K., and Regenstein J.M., “Changes in electrophoretic patterns of Gadoid and Non-Gadoid fish muscle during frozen storage”, J. Food Sci., 54(4): 819-823 (1989).
27. Türköz Y., Arslan A., Gönülalan Z., ve İleri T., “Değişik protein ekstraksiyon yöntemleri kullanarak balık türlerinin elektroforetik ayırımı, dondurarak saklama ve ısı işleminin kas proteinlerine etkisinin incelenmesi”, F.Ü. Sağlık Bil. Derg., 14(1): 31-38 (2000).
28. Pineiro C, Vazquez J, Marina AI, Barros-Velazquez J, Gallardo JM. “Characterization and partial sequencing of species-specific sarcoplasmic polypeptides from commercial hake species by mass spectrometry following two-dimensional electrophoresis”, Electrophoresis. 22(8): 1545-52 (2001).