Abstract
Activity of carnosinase (CN1), the only dipeptidase with substrate specificity for carnosine or homocarnosine, varies greatly between individuals but increases clearly and significantly with age. Surprisingly, the lower CN1 activity in children is not reflected by differences in CN1 protein concentrations. CN1 is present in different allosteric conformations in children and adults since all sera obtained from children but not from adults were positive in ELISA and addition of DTT to the latter sera increased OD450 values. There was no quantitative difference in the amount of monomeric CN1 between children and adults. Further, CN1 activity was dose dependently inhibited by homocarnosine. Addition of 80 μM homocarnosine lowered V max for carnosine from 440 to 356 pmol/min/μg and increased K m from 175 to 210 μM. The estimated K i for homocarnosine was higher (240 μM). Homocarnosine inhibits carnosine degradation and high homocarnosine concentrations in cerebrospinal fluid (CSF) may explain the lower carnosine degradation in CSF compared to serum. Because CN1 is implicated in the susceptibility for diabetic nephropathy (DN), our findings may have clinical implications for the treatment of diabetic patients with a high risk to develop DN. Homocarnosine treatment can be expected to reduce CN1 activity toward carnosine, resulting in higher carnosine levels.
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References
Aydogan S, Yapislar H, Artis S et al (2008) Impaired erythrocytes deformability in H82)O(2)-induced oxidative stress: protective effect of l-carnosine. Clin Hemorheaol Microcirc 39:93–98
Baguet A, Reyngoudt H, Pottier A et al (2009) Carnosine loading and washout in human skeletal muscles. J Appl Physiol 106:837–842
Balion CM, Benson C, Raina PS et al (2007) Brain type carnosinase in dementia: a pilot study. BMC Neurol 7:38
Bando K, Shimotsuji T, Toyoshima H (1984) Fluorometric assay of human serum carnosinase activity in normal children, adults and patients with myopathy. Ann Clin Biochem 21:510–514
Baran EJ (2000) Metal complexes of carnosine. Biochemistry (Mosc) 65:757–765
Baslow MH (2009) A novel key-lock mechanism for inactivating amino acid neurotransmitters during transit across extracellular space. Amino Acids (epub ahead of print)
Bauer K (2005) Carnosine and homocarnosine, the forgotten, enigmatic peptides of the brain. Neurochem Res 30:1339–1345
Boldyrev AA (1993) Does carnosine possess direct antioxidant activity? Int J Biochem 25:1101–1107
Boldyrev AA, Severin SE (1990) The histidine-containing dipeptides, carnosine and anserine: distribution, properties and biological significance. Adv Enzyme Regul 30:175–194
Boldyrev AA, Koldobski A, Kurella E et al (1993) Natural histidine-containing dipeptide carnosine as a potent hydrophilic antioxidant with membrane stabilizing function. A biomedical aspect. Mol Chem Neuropathol 19:185–192
Crush KG (1970) Carnosine and related substances in animal tissues. Comp Biochem Physiol 34:3–30
Duane P, Peters TJ (1988) Serum carnosinase activities in patients with alcoholic chronic skeletal muscle myopathy. Clin Sci (Lond) 75:185–190
Dunnett M, Harris RC, Dunnett CE et al (2002) Plasma carnosine concentration: diurnal variation and effects of age, exercise and muscle damage. Equine Vet J Suppl 34:283–287
Gjessing LR, Lunde HA, Morkid L et al (1990) Inborn errors of carnosine and homocarnosine metabolism. J Neural Transm Suppl 29:91–106
Hipkiss AR (1998) Carnosine, a protective, anti-ageing peptide? Int J Biochem Cell Biol 30:863–868
Jansen EW, Gibson KM, Shigematsu Y et al (2006) A novel, quantitative assay for homocarnosine in cerebrospinal fluid using stable-isotope dilution liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 830:196–200
Janssen B, Hohenadel D, Brinkkoetter P et al (2005) Carnosine as a protective factor in diabetic nephropathy. Association with a leucine repeat of the carnosinase gene CNDP1. Diabetes 54:2320–2327
Kendrick IP, Harris RC, Kim HJ et al (2008) The effects of 10 weeks of resistance training combined with beta-alanine supplementation on whole body strength, force production, muscular endurance and body composition. Amino Acids 34:547–554
Kim HJ (2009) Comparison of the carnosine and taurine contents of vastus lateralis of elderly Korean males, with impaired glucose tolerance, and young elite Korean swimmers. Amino Acids 36:359–363
Lenney JF, Georg RP, Weiss AM et al (1982) Human serum carnosinase: characterization, distinction from cellular carnosinase, and actvitation by cadmium. Clin Chim Acta 123:221–231
Lenney JF, Peppers SC, Kucera-Orallo CM et al (1985) Characterization of human tissue carnosinsase. Biochem J 228:653–660
Margolis FL, Grillo M, Brown CE et al (1979) Enzymatic and immunological evidence for two forms of carnosinase in the mouse. Biochim Biophys Acta 570:311–323
McFarland GA, Holliday R (1994) Retardation of the senescence of cultured human diploid fibroblasts by carnosine. Exp Cell Res 212:167–175
Min J, Senut MC, Rajanikant K (2008) Differential neuroprotective effects of carnosine, anserine and N-acetyl carnosine against permanent focal ischemia. J Neurosci Res 86:2984–2991
Pegova A, Abe H, Boldyrev A (2000) Hydrolysis of carnosine and related compounds by mammalian carnosinases. Comp Biochem Physiol B Biochem Mol Biol 127:443–446
Perry TL, Hansen S, Stedman D et al (1968) Homocarnosine in human cerebrospinal fluid: an age-dependent phenomenon. J Neurochem 15:1203–1206
Perry TL, Hansen S, Kennedy J (1974) CSF amino acids and plasma–CSF amino acid ratios in adults. J Neurochem 24:587–589
Quinn PJ, Boldyrev AA, Formazuyk VE (1992) Carnosine: its properties, functions and potential therapeutic applications. Mol Aspects Med 13:379–444
Schönherr J (2002) Analysis of products of animal origin in feeds by determination of carnosine and related dipeptides by high-performance liquid chromatography. J Agric Food Chem 27:1945–1950
Tabakman R, Lazarovici P, Kohen R (2002) Neuroprotective effects of carnosine and homocarnosine on pheochromocytoma PC12 cells exposed to ischemia. J Neurosci Res 68:463–469
Teufel M, Saudek V, Ledig J-P et al (2003) Sequence identification and characterization of human carnosinase and a closely related non-specific dipeptidase. JBC 278:6521–6531
Trombley PQ, Horning MS, Blakemore LJ (2000) Interactions between carnosine and zinc and copper: implications for neuromodulation and neuroprotection. Biochemistry (Mosc) 65:807–816
Vistoli G, Pedretti A, Cattaneo M et al (2006) Homology modelling of human serum carnosinase, a potential medicinal target, and MD simulation of its allosteric activation by citrate. J Med Chem 49(11):3269–3277
Wassif WS, Sherwood RA, Amir A (1994) Serum carnosinase activities in central nervous system disorders. Clin Chim Acta 225:57–64
Willi SM, Zhang Y, Hill JB et al (1997) A deletion in the long arm of chromosome 18 in a child with serum carnosinase deficiency. Pediatr Res 41:210–213
Acknowledgments
We thank Prof. K. Michael Gibson for comments and critical reading of the manuscript. Part of this study was supported by the EU-funded specific-target project PREDICTIONS on the identification of risk factors for the development of diabetic nephropathy as well as grants by the Deutsche Forschungsgemeinschaft to M. Mack and J Zschocke (Ma2510/3-1 and Zs17/5-1).
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Peters, V., Kebbewar, M., Jansen, E.W. et al. Relevance of allosteric conformations and homocarnosine concentration on carnosinase activity. Amino Acids 38, 1607–1615 (2010). https://doi.org/10.1007/s00726-009-0367-z
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DOI: https://doi.org/10.1007/s00726-009-0367-z