Purification and properties of human serum carnosinase.
Carnosinase from human plasma was purified 18,000-fold to apparent homogeneity in a four step procedure. The dipeptidase was partially inactivated during DEAE-cellulose chromatography; however, it reactivated slowly when concentrated and stored at 4 degrees C. In the second purification step, hydroxylapatite column chromatography, two forms of the enzyme were separated from one another. Human serum carnosinase was found to be a glycoprotein with a pI of 4.4 and a subunit Mr of 75,000; the active enzyme was a dimer, the two subunits being connected by one or more disulfide bonds. The enzyme was especially active in hydrolyzing carnosine and anserine, preferring dipeptides with histidine in the C-terminal position. In most human tissues, the concentration of serum carnosinase was proportional to the percentage of trapped blood in the sample. However, the brain contained about 9 times more enzyme than expected, based on the amount of trapped blood present. The physiological function of this enzyme seems to be the hydrolysis of homocarnosine in the brain and the splitting of carnosine and anserine in the blood stream. Six higher primates were found to have serum carnosinase. Twelve nonprimate mammals were tested; all were lacking the serum enzyme except for the Golden hamster, which had very high concentrations of a carnosinase having somewhat different properties than the higher primate enzyme.
Jackson
M C
MC
Pharmacology Department, School of Medicine, University of Hawaii, Honolulu 96822.
Kucera
C M
CM
Lenney
J F
JF
D000367:Age Factors; D000595:Amino Acid Sequence; D000818:Animals; D000861:Anserine / Q000378:metabolism; D001921:Brain / Q000201:enzymology*; D002336:Carnosine / Q000031:analogs & derivatives / Q000378:metabolism; D002848:Chromatography, DEAE-Cellulose; D006224:Cricetinae; D004150:Dipeptidases / Q000097:blood* / Q000134:cerebrospinal fluid / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000032:analysis; D000882:Haplorhini / Q000378:metabolism; D006801:Humans; D007525:Isoelectric Focusing; D008297:Male; D008647:Mesocricetus / Q000378:metabolism; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D013045:Species Specificity
D006023:Glycoproteins; C005038:homocarnosine; D002336:Carnosine; D004150:Dipeptidases; C019673:aminoacyl-histidine dipeptidase; D000861:Anserine