Characterization of two novel proteins, NgRH1 and NgRH2, structurally and biochemically homologous to the Nogo-66 receptor.
Nogo-66 receptor (NgR) has recently been identified as the neuronal receptor of the myelin-associated proteins Nogo-A, oligodendrocyte protein (OMgp) and myelin-associated glycoprotein (MAG), and mediates inhibition of axonal regeneration both in vitro and in vivo. Through database searches, we have identified two novel proteins (NgRH1 and NgRH2) that turned out to be homologous in their primary structures, biochemical properties and expression patterns to NgR. Like NgR, the homologues contain eight leucine-rich repeats (LRR) flanked by a leucine-rich repeat C-terminus (LRRCT) and a leucine-rich repeat N-terminus (LRRNT), and also have a C-terminal GPI signal sequence. Northern blot analysis showed predominant expression of NgRH1 and NgRH2 mRNA in the brain. In situ hybridization and immunohistochemistry on rat brain slices revealed neuronal expression of the genes. NgRH1 and NgRH2 were detected on the cell surface of recombinant cell lines as N-glycosylated GPI anchored proteins and, consistent with other GPI anchored proteins, were localized within the lipid rafts of cellular membranes. In addition, an N-terminal proteolytic fragment of NgR comprising the majority of the ectodomain was found to be constitutively secreted from cells. Our data indicate that NgR, NgRH1 and NgRH2 constitute a novel receptor protein family, which may play related roles within the CNS.
Pignot
V
V
Novartis Pharma Research, Basel, Switzerland.
Hein
A E
AE
Barske
C
C
Wiessner
C
C
Walmsley
A R
AR
Kaupmann
K
K
Mayeur
H
H
Sommer
B
B
Mir
A K
AK
Frentzel
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D058851:GPI-Linked Proteins; D015870:Gene Expression; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism; D008969:Molecular Sequence Data; D009185:Myelin Proteins / Q000096:biosynthesis / Q000235:genetics*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D000070816:Nogo Receptor 1; D000070817:Nogo Receptor 2; D000070156:Nogo Receptors; D009928:Organ Specificity; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D058851:GPI-Linked Proteins; D017261:Glycosylphosphatidylinositols; D009185:Myelin Proteins; D000070816:Nogo Receptor 1; D000070817:Nogo Receptor 2; D000070156:Nogo Receptors; D012333:RNA, Messenger; C486580:RTN4R protein, human; C474328:RTN4RL1 protein, human; C474327:RTN4RL2 protein, human; D011956:Receptors, Cell Surface; C486581:Rtn4r protein, rat