title: Stabilization of human prostatic acid phosphatase by coupling with chondroitin sulfate.
abstract: Human prostatic acid phosphatase (PAP) (EC 3.1.3.2) was covalently linked to chondroitin sulfate A from whale cartilage. In order to bind the protein amino groups with the preactivated carboxyl groups of chondroitin sulfate, 1-ethyl-3-(3'-dimethylaminepropyl)carbodiimide and N-hydroxysulfosuccinimide were used as coupling agents. The product was soluble and enzymatically active. The activity was on average 25% higher than that of the free enzyme. The product was heterogeneous in respect to charge and Mr (50-1500) kDa, as determined by chromatography on Sephacryl S 300 and polyacrylamide gel electrophoresis. The resulting polymers contained covalently bound chondroitin sulfate, as shown by the biotin-avidin test. The modified enzyme is more resistant against various denaturing agents, e.g., urea, ethanol, and heat. Thus covalent modification of PAP by cross-linking to chondroitin sulfate could be the preferred method for stabilization of its biological activity.
authors.affiliation: Institute of Medical Biochemistry, Medical Academy, Kraków, Poland.
authors.forename: E
authors.initials: E
authors.lastname: Luchter-Wasylewska
authors.forename: J
authors.initials: J
authors.lastname: Dulińska
authors.forename: W S
authors.initials: WS
authors.lastname: Ostrowski
authors.forename: V P
authors.initials: VP
authors.lastname: Torchilin
authors.forename: V S
authors.initials: VS
authors.lastname: Trubetskoy
mesh_terms: D000135:Acid Phosphatase / Q000378:metabolism*; D002809:Chondroitin Sulfates / Q000378:metabolism*; D003432:Cross-Linking Reagents; D004795:Enzyme Stability; D006801:Humans; D008297:Male; D015394:Molecular Structure; D011467:Prostate / Q000201:enzymology*; D011485:Protein Binding; D013056:Spectrophotometry, Ultraviolet
chemical_list: D003432:Cross-Linking Reagents; D002809:Chondroitin Sulfates; D000135:Acid Phosphatase