C-mannosylation of human hyaluronidase 1: Possible roles for secretion and enzymatic activity

Goto,Yuki; Niwa,Yuki; Suzuki,Takehiro; Dohmae,Naoshi; Umezawa,Kazuo; Simizu,Siro

doi:10.3892/ijo.2014.2438

C-mannosylation of human hyaluronidase 1: Possible roles for secretion and enzymatic activity

Authors:
- Yuki Goto
- Yuki Niwa
- Takehiro Suzuki
- Naoshi Dohmae
- Kazuo Umezawa
- Siro Simizu
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Affiliations: Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama, Japan, Global Research Cluster, RIKEN, Japan, Department of Molecular Target Medicine Screening, Aichi Medical University School of Medicine, Japan
Published online on: May 12, 2014 https://doi.org/10.3892/ijo.2014.2438
Pages: 344-350

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Abstract

Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. In the protein glycosylation, C-mannosylation was first identified in ribonuclease 2, and some proteins have been reported to be C-mannosylated; however, effects of its modifications for target proteins remain unclear. Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp130 and Trp321. In this study, we examined whether HYAL1 is C-mannosylated or not, and the effect of C-mannosylation on HYAL1. Using mass spectrometry, we first demonstrated that intracellular HYAL1 is C-mannosylated at Trp130 but not at Trp321. Surprisingly, although HYAL1 was secreted into conditioned medium and it possessed enzymatic activity, secreted HYAL1 was not C-mannosylated. Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp130 changed conformation of the catalytic active site, and faced Glu131 in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity.

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