Biochemical Study of Dipeptidyl Peptidase-4 in Autistic's Patients

Abstract

The research includes estimation of autistics' serum dipeptidyl peptidase-4 (DPP-4) activity. The results indicated a significant (p≤0.05) decrease in (DPP-4) activity of autism spectrum disorder groups, 57.85, 42.86, 36.47 µmol/L respectively compared to control group. By statistical analysis, the study revealed a significant (p≤0.05) relationship between DPP-4 activity with gastrointestinal disorder on one hand and with various inflammation incidence on the other hand. Partial purification of DPP-4 from serum of normal person age 14 years in Mosul city was done. Gel filtration of dialysate precipitate produced by 50% ammonium sulphate saturation has given two major proteinous components. One of them ( peak A) possesses a high DPP-4 activity using sephedex G-100. The apparent molecular weight of the isolated DPP-4 was 176.6 KD. Then SDS-PAGE was performed. HPLC revealed a single peak A' at retention time 5.829 min by application the top of peak A which was isolated from gel filtration. Maximum activity of DPP-4 was obtained using 0.1 M Tris-HCl buffer at pH 8, 40°C, 4 mM of gly-pro-p-nitroanilide hydrochloride as a substrate. Maximum velocity (Vmax) was 50 μM according to Line Weaver-Burk plot while Michaelis–Menten constant (Km) was 0.5 mM. Mercuric chloride and strontium chloride hexahydrate at 5 mM revealed maximum inhibitory effect of DPP4 activity by 30.2% and 42.9% respectively.

Keywords

• Dipeptidyl peptidase 4,Autism spectrum disorder,Gel filtration,Electrophoresis,High performance liquid chromatography

References

1. Al-Hayaly, M. Z. (2010). Estimation of lead level among children with pervasive developmental disorders. M.Sc. Thesis. College of Nursing. University of Mosul. Davy, A.; Thomsen, K.K.; Juliano, M.A.; Alves, L.C.; Svendsen, I.; Simpson, D.J. (2000). Purification and characterization of barley dipeptidyl peptidase IV. Plant Physiology, 122, 425-431. Detel, D.; Pugel, E.; Baticic, L.; Budjevic, P.S.; Varljen J. (2012). Development and resolution of colitis in mice with target deletion of dipeptidylpeptidase IV. Experimental physiology DOI:10.1113/expphysiol 2011.061143, 1-11. dipeptide substrate specificity. The Journal of Biological Chemistry. 281, 7437-7444. Duke-Cohan, J.; Moriamoto, C.; Schlossman, S.F. (2001). Form of dipeptidylpeptidase IV (CD26) found in human serum. United State Patents, 6, 325-989BI. Durinx, C.; Lambeir, A.; Bosmans, E.; Falmagne, J.; Berghmans, R.; Heamers, A.; Schorp, S.; Meester, I.D. (2000). Characterization of dipeptidylpeptidase activity in serum soluble CD26/ dipeptidylpeptidase IV is responsible for the release of X-Pro dipeptides. Eur. J. Biochem., 267: 5608-5613 Edosada, C.Y.; Quan, C.; Weismann, C.; Tran, T.; Sutherlin, D.; Reynolds, M.; Elliott, J.M.; Raab, H.; Fairbrother, W.; Beni, B.(2006). Enzyme catalysis and regulation: Selective inhibition of fibroblast activation protein protease based on dipeptide substrate specificity. The J. Bio. Chem., 281, 7437-7444. Evans, T. A.; Siedlak, S. L.; Lu, L.; Fu, X.; Wang, Z.; McGnnis, W. R.; Fakhoury, E.; Castellani, R. J.; Hazen, S. L.;Walsh, W. J.; Lewis, A. T.; Salomon, R. G.; Smith, M. A.; Perry, G.; Zhu, X. (2008). The autistic phenotype exhibits a remarkably localized modification of brain protein by products of free radical-induced lipid oxidation. American Journal of Biotechnology and Biochemistry, 4(2): 61-72. Gomez, H.; Chappe, M.; Valiente, P.A.; Pons, T.; Chavez, M.A.; Charli, J.L., Pascual, I. (2013). Effect of zinc and calcium ions on the rat kidney membrane – bound form of dipeptidyl peptidase IV. J. Biosci, 38(3), 1-9. Iwaki-Egawa, S.; Watanabe, Y.; Kikuya, Y.; Fujimoto, Y. (1998). Dipeptidylpeptidase IV from human serum : purification, characterization and N-terminal amino acid sequence. J. Biochem. 124, 428-433. King, M.W.(2012). MedicalBiochemistryPage. /themedicalbiochemistrypage.org/ Kreisel, W.; Heussner, R.; Volk, B.; Buchsel, R.; Reutter, W.; Gerok, W. (1982). Identification of the 110000 molecular weightglycoprotein isolated from rat liver plasma membrane as dipeptidyl amino peptidase IV. FEBS Lett., 147, 85-88. Lad, M. M. (2007). Biomedical approach for autism (Basics). Defeat Autism Now(DAN) model. Panchapakesan, U.; Gross,S.; Komala, M. G.; Pegg, K.; Pollak, C. A. (2013). DPP-4 Inhibition in human kidney proximal tubular cells-Reno protection in diabetic nephropathy?. Journal of Diabetes & Metabolism. S9. Pereira, D.A.; Gomez, L.; El-Cheikh, M.C.; Borojevic, R. (2003). Dipeptidyl peptidase IV(CD26) activity in the hematopoietic system: differences between the membrane-anchored and the released enzyme activity. Brazilian J. Medical and Bio. Research, 36, 567-578. Prabavathy, N.; Vijayakumari, M.; Minil, M.; Sathiyaraj, U.; Kavimani, S. (2011). Lingaleptin–A novel DPP-4 inhibitor. International Journal of Pharma & Bio Sciences, 2(1), 438-442. Randolph-Gips, M. M. and Srinivasan, P. (2012). Modeling autism: A systematic biology approach. Journal of clinical Bioinformatics, 2(17):1-32. Robyt, F.J.; White, J.B. (1987). Biochemical. Theory and Practice Booke .California, U.S.A. p. 33. Rose, S.; Melnky, R.; Trusy, T. A.; Pavliv, O.; Serdel, L.; Li, J., Todd, N.; James, S. J. (2012). Intracellular and extracellular redox status and free radicals generation in primary immune cells from children with autism. Autism research and treatment.1-10. Schacterle, G.R.; Pollack. (1973). A simplified method for the quantitative assay of small amount of protein in biological materials. Anal. Biochem., 51, 654-655. Sanz, Y. and Toldra, F. (2001). Purification and characterization of an X-prolyl- dipeptidyl peptidase from Lactobacillus sakei. Applied and environmental microbiology, 67(4):1815-1820. Shattock, P. and Whitely, P. (2002). Biochemical aspects in autism spectrum disorders:Updating the opioid-excess theory and presenting new opportunities for biomedical intervention. Expert Opinion. Ther. Targets, 6(2):175-183. Shaw, W. (2008). Biological Treatment For Autism and PDD – causes and biomedical therapies for autism and PDD-Abnormalities of the digestive system. 3rd ed.USA.pp 77-78. Shibuya-Saruta, H.; Kasahara, Y.; Hashimoto, Y. (1996). Human serum dipeptidylpeptidase IV(DPPIV) and its unique properties. J. Clin. Lab. Anal, 10(6), 435-40. Vanderheyden, M.; Batunek, J.; Coethals, M.; Verstraken, S.; Lambeir, A.M.; De Meester, I., Scharpe, S. (2009). Dipeptidylpeptidase IV and B type nutriuretic peptide. Clin. Chem. Lab. Med., 47(3), 248-252. Volkmar, F. R. and Pauls, D. (2003). Autism. The Lancet, 362(9390):1133-1141.