Defectively N-glycosylated and non-O-glycosylated aminopeptidase N (CD13) is normally expressed at the cell surface and has full enzymatic activity

K Norén; Gert Helge Hansen; H Clausen; Ove Norén; H Sjöström; L K Vogel

doi:10.1006/excr.1996.3455

Defectively N-glycosylated and non-O-glycosylated aminopeptidase N (CD13) is normally expressed at the cell surface and has full enzymatic activity

K Norén, Gert Helge Hansen, H Clausen, Ove Norén, H Sjöström, L K Vogel

Institut for Cellulær og Molekylær Medicin

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

14 Citationer (Scopus)

Abstract

Udgivelsesdato: 1997-Feb-25

Originalsprog	Engelsk
Tidsskrift	Experimental Cell Research
Vol/bind	231
Udgave nummer	1
Sider (fra-til)	112-8
Antal sider	6
ISSN	0014-4827
DOI	https://doi.org/10.1006/excr.1996.3455
Status	Udgivet - 1997

Bibliografisk note

Keywords: Animals; Antigens, CD13; Biological Transport; Cell Line; Cell Membrane; DNA, Complementary; Glycosylation; Humans; Polysaccharides; Swine; Transfection

Adgang til dokumentet

10.1006/excr.1996.3455

Citationsformater

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title = "Defectively N-glycosylated and non-O-glycosylated aminopeptidase N (CD13) is normally expressed at the cell surface and has full enzymatic activity",

abstract = "In order to study the effects of the absence of O-glycosylation and modifications of N-glycosylation on a class II membrane protein, pig and human aminopeptidase N (CD13) were stably expressed in the ldl(D) cell line. This cell line carries a UDP-Gal/UDP-GalNAc-epimerase deficiency which blocks the conversion of glucose into galactose derivatives. Thus it is possible in the ldl(D) cell line to selectively block O-glycosylation by the omission of N-acetylgalactoseamine from the culture medium and to alter N-glycosylation by the omission of galactose. In this way selectively altered glycosylated forms of the glycoprotein aminopeptidase N can be synthesized and the effects of altered glycosylation can be studied. It is demonstrated that aminopeptidase N carries {"}mucin-type{"} O-glycans and that this is predominantly located in the stalk, which connects the catalytic headgroup to the membrane anchor. Normally glycosylated aminopeptidase N is present in the plasma membrane of the ldl(D) cells. This is also the case for the non-O-glycosylated and defectively N-glycosylated forms. This is in line with the finding that the intracellular transport APN is unaffected by the absence of O-glycosylation or by changes in N-glycosylation as the various glycosylated forms of aminopeptidase N are normally converted from the high-mannose form to the complex glycosylated form. Enzymatic activity is not influenced by the changes in glycosylation.",

author = "K Nor{\'e}n and Hansen, {Gert Helge} and H Clausen and Ove Nor{\'e}n and H Sj{\"o}str{\"o}m and Vogel, {L K}",

note = "Keywords: Animals; Antigens, CD13; Biological Transport; Cell Line; Cell Membrane; DNA, Complementary; Glycosylation; Humans; Polysaccharides; Swine; Transfection",

year = "1997",

doi = "10.1006/excr.1996.3455",

language = "English",

volume = "231",

pages = "112--8",

journal = "Experimental Cell Research",

issn = "0014-4827",

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number = "1",

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TY - JOUR

T1 - Defectively N-glycosylated and non-O-glycosylated aminopeptidase N (CD13) is normally expressed at the cell surface and has full enzymatic activity

AU - Norén, K

AU - Hansen, Gert Helge

AU - Clausen, H

AU - Norén, Ove

AU - Sjöström, H

AU - Vogel, L K

N1 - Keywords: Animals; Antigens, CD13; Biological Transport; Cell Line; Cell Membrane; DNA, Complementary; Glycosylation; Humans; Polysaccharides; Swine; Transfection

PY - 1997

Y1 - 1997

N2 - In order to study the effects of the absence of O-glycosylation and modifications of N-glycosylation on a class II membrane protein, pig and human aminopeptidase N (CD13) were stably expressed in the ldl(D) cell line. This cell line carries a UDP-Gal/UDP-GalNAc-epimerase deficiency which blocks the conversion of glucose into galactose derivatives. Thus it is possible in the ldl(D) cell line to selectively block O-glycosylation by the omission of N-acetylgalactoseamine from the culture medium and to alter N-glycosylation by the omission of galactose. In this way selectively altered glycosylated forms of the glycoprotein aminopeptidase N can be synthesized and the effects of altered glycosylation can be studied. It is demonstrated that aminopeptidase N carries "mucin-type" O-glycans and that this is predominantly located in the stalk, which connects the catalytic headgroup to the membrane anchor. Normally glycosylated aminopeptidase N is present in the plasma membrane of the ldl(D) cells. This is also the case for the non-O-glycosylated and defectively N-glycosylated forms. This is in line with the finding that the intracellular transport APN is unaffected by the absence of O-glycosylation or by changes in N-glycosylation as the various glycosylated forms of aminopeptidase N are normally converted from the high-mannose form to the complex glycosylated form. Enzymatic activity is not influenced by the changes in glycosylation.

AB - In order to study the effects of the absence of O-glycosylation and modifications of N-glycosylation on a class II membrane protein, pig and human aminopeptidase N (CD13) were stably expressed in the ldl(D) cell line. This cell line carries a UDP-Gal/UDP-GalNAc-epimerase deficiency which blocks the conversion of glucose into galactose derivatives. Thus it is possible in the ldl(D) cell line to selectively block O-glycosylation by the omission of N-acetylgalactoseamine from the culture medium and to alter N-glycosylation by the omission of galactose. In this way selectively altered glycosylated forms of the glycoprotein aminopeptidase N can be synthesized and the effects of altered glycosylation can be studied. It is demonstrated that aminopeptidase N carries "mucin-type" O-glycans and that this is predominantly located in the stalk, which connects the catalytic headgroup to the membrane anchor. Normally glycosylated aminopeptidase N is present in the plasma membrane of the ldl(D) cells. This is also the case for the non-O-glycosylated and defectively N-glycosylated forms. This is in line with the finding that the intracellular transport APN is unaffected by the absence of O-glycosylation or by changes in N-glycosylation as the various glycosylated forms of aminopeptidase N are normally converted from the high-mannose form to the complex glycosylated form. Enzymatic activity is not influenced by the changes in glycosylation.

U2 - 10.1006/excr.1996.3455

DO - 10.1006/excr.1996.3455

M3 - Journal article

C2 - 9056417

SN - 0014-4827

VL - 231

SP - 112

EP - 118

JO - Experimental Cell Research

JF - Experimental Cell Research

IS - 1

ER -