+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Characterization of two carnosine-degrading enzymes from rat brain. Partial purification and characterization of a carnosinase and a beta-alanyl-arginine hydrolase

Characterization of two carnosine-degrading enzymes from rat brain. Partial purification and characterization of a carnosinase and a beta-alanyl-arginine hydrolase

European Journal of Biochemistry 160(3): 605-613

From rat brain extracts, two carnosine-degrading enzymes have been identified and partially purified by ion-exchange chromatography, hydrophobic interaction chromatography on phenyl-Sepharose CL-4B and gel filtration. These enzymes exhibit distinct differences in their chemical characteristics and substrate specificities. One enzyme, designated carnosinase, preferentially hydrolyzes carnosine and exhibits a low Km value (0.02 mM) towards this substrate. Carnosinase also degrades anserine but not homocarnosine or homoanserine. The other carnosine-degrading enzyme hydrolyzes beta Ala-Arg considerably faster than carnosine and, therefore, has been tentatively designated beta Ala-Arg hydrolase. This enzyme exhibits high Km values with carnosine (Km = 25 mM) and beta Ala-Arg (Km = 2 mM). Homocarnosine and gamma-aminobutyryl-arginine are not degraded by beta Ala-Arg hydrolase. Neither enzyme is inhibited by agents reactive on activated hydroxyl groups, such as diisopropyl fluorophosphate, and also not by a variety of peptidase inhibitors of microbial origin or from other sources. Carnosinase is also not inhibited by bestatin but beta Ala-Arg hydrolase, although not an aminopeptidase, is strongly inhibited by this aminopeptidase inhibitor (IC50 = 50 nM). While carnosinase is strongly inhibited by thiol-reducing agents such as dithioerythritol and 2-mercaptoethanol, beta Ala-Arg hydrolase is stabilized and activated by these substances. Both enzymes are strongly inhibited by metal-chelating agents. Carnosinase, however, is not dependent on exogeneously added metal ions and is strongly inhibited by Mn2+ as well as by heavy metal ions. In contrast, beta Ala-Arg hydrolase requires Mn2+ ions for full enzymatic activity. Based on these differences, selective incubation conditions could be evaluated in order to determine specifically both enzyme activities in crude tissue extracts. In rat, both enzymes are present in all tissues tested, except skeletal muscles, but considerable differences in their relative distribution among different tissues are also observed.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 039526909

Download citation: RISBibTeXText

PMID: 3780724

Related references

Characterization of two carnosine-degrading enzymes from rat brain : Partial purification and characterization of a carnosinase and a β-alanyl-arginine hydrolase. Febs Journal 160(3): 605-613, 1986

Separation of a new alpha n benzoyl arginine beta naphthylamide hydrolase from cathepsin b 1 ec purification characterization and properties of both enzymes from rabbit lung/. Journal of Biological Chemistry 253(12): 4319-4327, 1978

A new sensitive method for the determination of serum carnosinase activity using l-carnosine-[I-14C] beta-alanyl as substrate. Clinica Chimica Acta; International Journal of Clinical Chemistry 29(2): 243-248, 1970

Partial purification and characterization of enzymes degrading trehalose in mycobacteria. Annales Universitatis Mariae Curie-Sklodowska. Sectio D: Medicina 54: 285-289, 1999

Carnosine-anserine synthetase of muscle. 4. Partial purification of the enzyme and further studies of beta-alanyl peptide synthesis. Biochemical Journal 77(3): 575-581, 1960

Purification and partial characterization of an arginine ester hydrolase from the venom of the bushmaster snake, Lachesis muta noctivaga. Toxicon 23(4): 707-718, 1985

Isolation of poly(beta-L-malic acid)-degrading bacteria and purification and characterization of the PMA hydrolase from Comamonas acidovorans strain 7789. Fems Microbiology Letters 173(2): 365-372, 1999

A comparative study of ribonuclease from two winter barley cultivars based upon separation, purification and partial characterization of RNA-degrading enzymes. Dissertation Abstracts International, B 36(9): 4459B, 1976

Separation of a new alpha-N-benzoylarginine-beta-naphthylamide hydrolase from cathepsin B1. Purification, characterization, and properties of both enzymes from rabbit lung. Journal of Biological Chemistry 253(12): 4319-4326, 1978

Studies on xylan degrading enzymes part 1 purification and characterization of endo 1 4 beta xylanase ec from aspergillus niger strain 14. Biochimica et Biophysica Acta 484(1): 79-93, 1977

Partial purification and further characterization of the novel endoglucosaminidase from human serum that hydrolyses 4-methylumbelliferyl-N-acetyl-beta-D-chitotetraoside (MU-TACT hydrolase). International Journal of Biochemistry 26(12): 1369-1375, 1994

Biochemical characterization of the arginine degrading enzymes arginase and arginine deiminase and their effect on nitric oxide production. Medical Science Monitor 8(7): Br248-Br253, 2002

Production , purification and partial characterization of 1,4-beta-glucosidase enzymes from Sporotrichum pulverulentum. European Journal of Biochemistry 90(1): 191-198, 1978

Partial purification and characterization of enzymes associated with synthesis of beta lactam compounds in streptomyces clavuligerus. International Union Of Microbiological Societies 13th International Congress Of Microbiology; Boston, Mass , Usa, Aug 8-13, Xiv+182p American Society For Microbiology: Washington, D C , Usa Paper P121, 1982

Isolation and characterization of a novel simazine-degrading beta-proteobacterium and detection of genes encoding s-triazine-degrading enzymes. Pest Management Science 63(3): 261-268, 2007