Purification and properties of esterases characteristic of adult rat brain.
1. A method for the partial purification of an esterase fraction, present in the brain of the adult but not the newborn rat, is described. A 54-fold purification was achieved in three steps. 2. When subjected to starch-gel electrophoresis, the purified fraction resolved into three bands of esterase activity. Two of these bands migrate close together and faster than other esterases in the brain. These two esterases are inhibited by p-hydroxymercuribenzoate but not by di-isopropyl phosphorofluoridate. The third band is di-isopropyl phosphorofluoridate-sensitive and migrates just behind the two leading esterases. 3. After treatment with di-isopropyl phosphorofluoridate, to obviate the effects of the di-isopropyl phosphorofluoridate-sensitive esterase, the enzyme preparation hydrolyses alpha-naphthyl acetate, alpha-naphthyl propionate and alpha-naphthyl butyrate, but not cholesteryl acetate. The V(max.) for the naphthyl esters decreased with increase in chain length of the acyl group. The acetate ester is hydrolysed 34 times as fast as the butyrate and about seven times as fast as the propionate derivative. The K(m) values for these three esters, measured at pH7.2 and 37 degrees , are 2.8x10(-4)m, 3.1x10(-4)m and 7.3x10(-5)m for the acetate, propionate and butyrate derivatives respectively. 4. The Hofstee (1952) plots for the kinetic data show a single line, indicating that the two most-rapidly migrating esterases, although electrophoretically separable, are not kinetically distinguishable in the substrate ranges examined.
D000085:Acetates / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000201:enzymology*; D002087:Butyrates / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D004586:Electrophoresis; D004950:Esterases / Q000302:isolation & purification* / Q000378:metabolism*; D007531:Isoflurophate / Q000494:pharmacology; D007700:Kinetics; D008628:Mercury / Q000494:pharmacology; D009281:Naphthalenes / Q000378:metabolism; D009942:Organometallic Compounds / Q000494:pharmacology; D011422:Propionates / Q000378:metabolism; D051381:Rats
D000085:Acetates; D002087:Butyrates; D009281:Naphthalenes; D009942:Organometallic Compounds; D011422:Propionates; D007531:Isoflurophate; D002784:Cholesterol; D004950:Esterases; D008628:Mercury