The human CD10 lacking an N-glycan at Asn628 is deficient in surface expression and neutral endopeptidase activity

Ban Sato, Yohko U. Katagiri, Kazutoshi Iijima, Hiroyuki Yamada, Satsuki Ito, Nana Kawasaki, Hajime Okita, Junichiro Fujimoto, Nobutaka Kiyokawa

研究成果: Article査読

9 被引用数 (Scopus)

抄録

Background: CD10, also known as neprilysin or enkephalinase exhibiting neutral endopeptidase (NEP) activity, is expressed by B-lineage hematopoietic cells as well as a variety of cells from normal tissues. It cleaves peptides such as cytokines to act for terminating inflammatory responses. Although CD10 molecules of the human pre-B-cell line NALM-6 have 6 consensus N-glycosylation sites, three of them are known to be N-glycosylated by X-ray crystallography. Methods: In order to investigate the role of N-glycans in the full expression of NEP activity, we modified N-glycans by treatment of NALM6 cells with various glycosidases or alter each of the consensus N-glycosylation sites by generating site-directed mutagenesis and compared the NEP activities of the sugar-altered CD10 with those of intact CD10. Results: CD10 of the human B-cell line NALM-6 was dominantly localized in raft microdomains and heterogeneously N-glycosylated. Although neither desialylation nor further degalactosylation caused defective NEP activity, removal of only a small part of N-glycans by treatment with glycopeptidase F under non-denaturing conditions decreased NEP activity completely. All of the three consensus sites of CD10 in HEK293 cells introduced with wild type-CD10 were confirmed to be N-glycosylated. Surface expression of N-glycan at Asn628-deleted CD10 by HEK293 cells was greatly decreased as well as it lost entire NEP activities. Conclusions: N-glycosylation at Asn628 is essential not only for NEP activities, but also for surface expression. General significance: Quality control system does not allow dysfunctional ecto-type proteases to express on plasma membrane.

本文言語English
ページ(範囲)1715-1723
ページ数9
ジャーナルBiochimica et Biophysica Acta - General Subjects
1820
11
DOI
出版ステータスPublished - 2012 11月
外部発表はい

ASJC Scopus subject areas

  • 生物理学
  • 生化学
  • 分子生物学

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