title: N-glycosylation is required for secretion and enzymatic activity of human hyaluronidase1.
abstract: Hyaluronidase1 (HYAL1) is a hydrolytic enzyme that degrades hyaluronic acid (HA) and has three predicted N-glycosylation sites at Asn(99), Asn(216), and Asn(350). In this report, we show the functional significance of N-glycosylation on HYAL1 functions. Using mass spectrometry, we demonstrated that HYAL1 was N-glycosylated at the three asparagine residues. N-glycosylation of HYAL1 is important for secretion of HYAL1, as demonstrated by site-directed mutation. Moreover, a defect of N-glycosylation attenuated the enzymatic activity of HYAL1. Thus, HYAL1 is N-glycosylated at the three asparagine residues, and its secretion and enzymatic activity are regulated by N-glycosylation.
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Goto
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Niwa
authors.affiliation: Global Research Cluster, RIKEN, Wako 351-0198, Japan.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Suzuki
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan.
authors.forename: Shiho
authors.initials: S
authors.lastname: Uematsu
authors.affiliation: Global Research Cluster, RIKEN, Wako 351-0198, Japan.
authors.forename: Naoshi
authors.initials: N
authors.lastname: Dohmae
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan.
authors.forename: Siro
authors.initials: S
authors.lastname: Simizu
keywords: CBB, Coomassie Brilliant Blue; DMEM, Dulbecco’s modified Eagle’s medium; ECM, extracellular matrix; ER, endoplasmic reticulum; Enzymatic activity; HA, hyaluronic acid; HYAL1, hyaluronidase1; Hyaluronidase1; MALDI-TOF MS, matrix-assisted laser desorption ionization time of flight mass spectrometry; N-glycosylation; PBS, phosphate-buffered saline; SDS, sodium dodecyl sulfate; Secretion