Kurloff cell lysosomal arylsulphatases: presence of both cationic and highly anionic isoforms of the sole B class.
Following the previous ultrastructural demonstration of the presence of arylsulphatase (Asase) activities in Kurloff cells (KC) and of their quasi-exclusive localization in the Kurloff body (KB), this work investigates their biochemical and zymographic properties after extraction from purified KC suspensions. Using the discriminative inhibitory conditions of both the Baum or Lee-Vaupel and Conzelmann methods, nitrocatechol sulphate hydrolyzing enzymes of the KC were assumed to belong to the B class of the type II Asase alone. After electrophoretic separation under non-denaturing conditions in a 4-23% polyacrylamide gel, they were characterized by 55 kDa and 62 kDa zymographic bands. After isoelectric focusing, 'classical' cationic isoforms (pI 8.5) and two anionic isoforms (pI 4.4 and 4.6) were observed on zymograms. As expected for class B Asase, the different zymographic forms of KC Asase were only recovered in the unadsorbed fraction after anion-exchange chromatography on DEAE-cellulose column equilibrated with high ionic strength buffer. Their Km (2.1 mM), their optimum pH (5.8) and their inhibitions by sulfite, phosphate, sulphate and ascorbic acid as well as their slight stimulation by AgNO3 were also characteristic of this class of Asase. Finally, chondroitin-4-sulphate was shown to potentially be a physiological substrate for these lysosomal enzymes.
Taouji
S
S
Laboratoire d'Histologie et Biologie Cellulaire, UFR de Médecine, Université de Caen, France.
Buat
M L
ML
Izard
J
J
Landemore
G
G
D000818:Animals; D001192:Arylsulfatases / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D002848:Chromatography, DEAE-Cellulose; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006168:Guinea Pigs; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D007694:Killer Cells, Natural / Q000201:enzymology* / Q000648:ultrastructure; D007700:Kinetics; D008247:Lysosomes / Q000201:enzymology* / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D008970:Molecular Weight; D015388:Organelles / Q000201:enzymology* / Q000648:ultrastructure
D007527:Isoenzymes; D004958:Estradiol; D001192:Arylsulfatases