Studies on the charge isomers of arylsulfatase A.
Human liver arylsulfatase A was resolved into six fractions by narrow pH range preparative isoelectric focusing. Analytical isoelectric focusing revealed that most enzyme fractions were composed of two adjacent charge isomers. Nevertheless, there was considerable enrichment of charge species which allowed a comparative study of selected properties. Except for the most cationic fraction, neuraminidase treatment converted enzyme in all fractions to the three most cationic species. The most electronegative enzyme species had the highest molecular mass being made up of 64-kDa subunits. As electronegativity decreased, there was concomitant decrease in molecular mass and increase in complexity of subunit composition. Two subunits--61 and 55 kDa--prevailed with increasing proportions of the smaller unit with loss of electronegativity. There was also an increasing amount of a 26-kDa fraction which became a substantial component of the most cationic subfraction. Only enzyme in the two fractions containing the largest and most anionic species were taken up by cultured fibroblasts at higher efficiency than unfractionated enzyme. It is suggested that processing or maturation of arylsulfatase A incurs stepwise removal of charge groups and/or peptide segments leading to smaller, less-charged enzyme species.
Sarafian
T A
TA
Tsay
K K
KK
Jackson
W E
WE
Fluharty
A L
AL
Kihara
H
H
D002553:Cerebroside-Sulfatase / Q000032:analysis*; D004705:Endocytosis; D006801:Humans; D007525:Isoelectric Focusing; D008099:Liver / Q000201:enzymology; D009439:Neuraminidase / Q000494:pharmacology; D013429:Sulfatases / Q000032:analysis*
D013429:Sulfatases; D002553:Cerebroside-Sulfatase; D009439:Neuraminidase