title: Common G102S polymorphism in chitotriosidase differentially affects activity towards 4-methylumbelliferyl substrates.
abstract: Chitotriosidase (CHIT1) is a chitinase that is secreted by activated macrophages. Plasma chitotriosidase activity reflects the presence of lipid-laden macrophages in patients with Gaucher disease. CHIT1 activity can be conveniently measured using fluorogenic 4-methylumbelliferyl (4MU)-chitotrioside or 4MU-chitobioside as the substrate, however, nonsaturating concentrations have to be used because of apparent substrate inhibition. Saturating substrate concentrations can, however, be used with the newly designed substrate 4MU-deoxychitobioside. We studied the impact of a known polymorphism, G102S, on the catalytic properties of CHIT1. The G102S allele was found to be common in type I Gaucher disease patients in the Netherlands ( approximately 24% of alleles). The catalytic efficiency of recombinant Ser102 CHIT1 was approximately 70% that of wild-type Gly102 CHIT1 when measured with 4MU-chitotrioside at a nonsaturating concentration. However, the activity was normal with 4MU-deoxychitobioside as the substrate at saturating concentrations, consistent with predictions from molecular dynamics simulations. In conclusion, interpretation of CHIT1 activity measurements with 4MU-chitotrioside with respect to CHIT1 protein concentrations depends on the presence of Ser102 CHIT1 in an individual, complicating estimation of the body burden of storage macrophages. Use of the superior 4MU-deoxychitobioside substrate avoids such complications because activity towards this substrate under saturating conditions is not affected by the G102S substitution.
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Anton P
authors.initials: AP
authors.lastname: Bussink
authors.forename: Marri
authors.initials: M
authors.lastname: Verhoek
authors.forename: Jocelyne
authors.initials: J
authors.lastname: Vreede
authors.forename: Karen
authors.initials: K
authors.lastname: Ghauharali-van der Vlugt
authors.forename: Wilma E
authors.initials: WE
authors.lastname: Donker-Koopman
authors.forename: Richard R
authors.initials: RR
authors.lastname: Sprenger
authors.forename: Carla E
authors.initials: CE
authors.lastname: Hollak
authors.forename: Johannes M F G
authors.initials: JM
authors.lastname: Aerts
authors.forename: Rolf G
authors.initials: RG
authors.lastname: Boot
mesh_terms: D000483:Alleles; D019943:Amino Acid Substitution; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D005776:Gaucher Disease / Q000201:enzymology / Q000235:genetics; D005787:Gene Frequency; D006031:Glycosylation; D006596:Hexosaminidases / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000378:metabolism; D066298:In Vitro Techniques; D008958:Models, Molecular; D009426:Netherlands; D020641:Polymorphism, Single Nucleotide; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013816:Thermodynamics; D014312:Trisaccharides / Q000378:metabolism; D014468:Umbelliferones / Q000378:metabolism
chemical_list: C403643:4-methylumbelliferyl-chitotrioside; D017931:DNA Primers; D011994:Recombinant Proteins; D014312:Trisaccharides; D014468:Umbelliferones; D006923:Hymecromone; D006596:Hexosaminidases; C086400:chitotriosidase