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N‐glycosylation is required for efficient secretion of a novel human secreted glycoprotein, hPAP21
Author(s) -
Zhou Yu-Bo,
Liu Feng,
Zhu Zhi-Dong,
Zhu Hong,
Zhang Xin,
Wang Zhi-Qin,
Liu Jian-Hua,
Han Ze-Guang
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.09.039
Subject(s) - glycosylation , secretion , glycoprotein , chinese hamster ovary cell , pngase f , signal peptide , n linked glycosylation , biology , biochemistry , extracellular , secretory protein , transmembrane domain , immunoprecipitation , blot , microbiology and biotechnology , peptide sequence , gene , receptor , glycan
The present study reported the isolation and characterization of a novel human secreted protein, named as hPAP21 (human protease‐associated domain‐containing protein, 21 kDa), encoded by the hypothetical gene chromosome 2 open reading frame 7 ( C2orf7 ) that contains signal peptide in its N‐terminus, without transmembrane domain, except for PA domain in its middle. Western blotting assay indicated that the c‐Myc tagged hPAP21 could be secreted into culture medium in the transfected Chinese hamster ovary cells. However, the molecular weights, whatever intracellular (28 kDa) or extracellular (30 kDa) forms, are larger than that of the prediction. To define whether the glycosylation was important process for its secretion, endoglycosidase H (Endo H) and PNGase F (PNG F) were employed to evaluate the effect of glycosylation types on secretion of hPAP21. Interestingly, the extracellular forms were primarily sensitive to PNG F, not Endo H, implying that complex N‐glycosylation could be required for the secretion of hPAP21. Furthermore, N‐glycosylation of Asn171 was confirmed as potential crucial process for the secretory protein via site‐directed mutagenesis assay. All data will be contributed to the understanding of molecular functions of hPAP21.