The Uptake of Heparanase into Mast Cells Is Regulated by Its Enzymatic Activity to Degrade Heparan Sulfate

Shi, Jia; Onuki, Yoshiki; Kawanami, Fumiya; Miyagawa, Naoko; Iwasaki, Fumika; Tsuda, Haruna; Takahashi, Katsuhiko; Oku, Teruaki; Suzuki, Masato; Higashi, Kyohei; Adachi, Hayamitsu; Nishimura, Yoshio; Nakajima, Motowo; Irimura, Tatsuro; Higashi, Nobuaki

doi:10.3390/ijms25116281

Open AccessArticle

The Uptake of Heparanase into Mast Cells Is Regulated by Its Enzymatic Activity to Degrade Heparan Sulfate

by

Jia Shi

^1,†,

Yoshiki Onuki

^1,†,

Fumiya Kawanami

¹,

Naoko Miyagawa

¹,

Fumika Iwasaki

¹,

Haruna Tsuda

¹,

Katsuhiko Takahashi

¹,

Teruaki Oku

²

,

Masato Suzuki

³,

Kyohei Higashi

³

,

Hayamitsu Adachi

⁴,

Yoshio Nishimura

⁵,

Motowo Nakajima

⁶

,

Tatsuro Irimura

⁷

and

Nobuaki Higashi

^1,*

¹

Department of Biochemistry, Hoshi University School of Pharmacy, 2-4-41, Ebara, Shinagawa-ku 142-8501, Tokyo, Japan

²

Department of Microbiology, Hoshi University School of Pharmacy, 2-4-41, Ebara, Shinagawa-ku 142-8501, Tokyo, Japan

³

Department of Clinical and Analytical Biochemistry, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641, Yamazaki, Noda 278-8510, Chiba, Japan

⁴

Institute of Microbial Chemistry (BIKAKEN), 18-24, Miyamoto, Numazu 410-0301, Shizuoka, Japan

⁵

Institute of Microbial Chemistry (BIKAKEN), 3-14-23, Kamiosaki, Shinagawa-ku 141-0021, Tokyo, Japan

⁶

SBI Pharmaceuticals Co., Ltd., 1-6-1, Roppongi, Minato-ku 106-6019, Tokyo, Japan

⁷

Division of Glycobiologics, Juntendo University Graduate School of Medicine, 2-1-1, Hongo, Bunkyo-ku 113-8421, Tokyo, Japan

^*

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Int. J. Mol. Sci. 2024, 25(11), 6281; https://doi.org/10.3390/ijms25116281

Submission received: 8 May 2024 / Revised: 29 May 2024 / Accepted: 4 June 2024 / Published: 6 June 2024

(This article belongs to the Special Issue Heparin, Heparan Sulfate and Heparanase in Health and Disease)

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Abstract

Mast cells take up extracellular latent heparanase and store it in secretory granules. The present study examined whether the enzymatic activity of heparanase regulates its uptake efficiency. Recombinant mouse heparanase mimicking both the latent and mature forms (L-Hpse and M-Hpse, respectively) was internalized into mastocytoma MST cells, peritoneal cell-derived mast cells, and bone marrow-derived mast cells. The internalized amount of L-Hpse was significantly higher than that of M-Hpse. In MST cells, L-Hpse was continuously internalized for up to 8 h, while the uptake of M-Hpse was saturated after 2 h of incubation. L-Hpse and M-Hpse are similarly bound to the MST cell surface. The expression level of cell surface heparan sulfate was reduced in MST cells incubated with M-Hpse. The internalized amount of M-Hpse into mast cells was significantly increased in the presence of heparastatin (SF4), a small molecule heparanase inhibitor that does not affect the binding of heparanase to immobilized heparin. Enzymatically quiescent M-Hpse was prepared with a point mutation at Glu335. The internalized amount of mutated M-Hpse was significantly higher than that of wild-type M-Hpse but similar to that of wild-type and mutated L-Hpse. These results suggest that the enzymatic activity of heparanase negatively regulates the mast cell-mediated uptake of heparanase, possibly via the downregulation of cell surface heparan sulfate expression.

Keywords: endocytosis; heparanase; heparan sulfate degradation; heparastatin (SF4); heparin; mast cells; syndecan

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MDPI and ACS Style

Shi, J.; Onuki, Y.; Kawanami, F.; Miyagawa, N.; Iwasaki, F.; Tsuda, H.; Takahashi, K.; Oku, T.; Suzuki, M.; Higashi, K.; et al. The Uptake of Heparanase into Mast Cells Is Regulated by Its Enzymatic Activity to Degrade Heparan Sulfate. Int. J. Mol. Sci. 2024, 25, 6281. https://doi.org/10.3390/ijms25116281

AMA Style

Shi J, Onuki Y, Kawanami F, Miyagawa N, Iwasaki F, Tsuda H, Takahashi K, Oku T, Suzuki M, Higashi K, et al. The Uptake of Heparanase into Mast Cells Is Regulated by Its Enzymatic Activity to Degrade Heparan Sulfate. International Journal of Molecular Sciences. 2024; 25(11):6281. https://doi.org/10.3390/ijms25116281

Chicago/Turabian Style

Shi, Jia, Yoshiki Onuki, Fumiya Kawanami, Naoko Miyagawa, Fumika Iwasaki, Haruna Tsuda, Katsuhiko Takahashi, Teruaki Oku, Masato Suzuki, Kyohei Higashi, and et al. 2024. "The Uptake of Heparanase into Mast Cells Is Regulated by Its Enzymatic Activity to Degrade Heparan Sulfate" International Journal of Molecular Sciences 25, no. 11: 6281. https://doi.org/10.3390/ijms25116281

APA Style

Shi, J., Onuki, Y., Kawanami, F., Miyagawa, N., Iwasaki, F., Tsuda, H., Takahashi, K., Oku, T., Suzuki, M., Higashi, K., Adachi, H., Nishimura, Y., Nakajima, M., Irimura, T., & Higashi, N. (2024). The Uptake of Heparanase into Mast Cells Is Regulated by Its Enzymatic Activity to Degrade Heparan Sulfate. International Journal of Molecular Sciences, 25(11), 6281. https://doi.org/10.3390/ijms25116281

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The Uptake of Heparanase into Mast Cells Is Regulated by Its Enzymatic Activity to Degrade Heparan Sulfate

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