Role of the peripheral myelin protein 22 N-linked glycan in oligomer stability.
Peripheral myelin protein 22 (PMP22) is a 22-kDa glycoprotein containing a single N-linked carbohydrate moiety. This posttranslational modification is conserved in PMP22 across species and within members of the PMP22 gene family; however, the function of the oligosaccharide is not known. To study the role of the PMP22 carbohydrate, site-directed mutagenesis was used to alter the glycosylation consensus sequence and produce a glycosylation-deficient mutant protein. This modified PMP22 was expressed in primary Schwann cells (SCs), and the effect of the N-glycan on the turnover rate, oligomerization, and intracellular trafficking of PMP22 was determined. Our data show a slight decrease in turnover rate from a half-life of approximately 70 min for the wild-type (wt) protein to 100 min for the glycosylation mutant. Although the presence of glycosylation-deficient PMP22 oligomers could be detected in SCs, we observed a decrease in oligomer stability compared with the wt oligomers. Both wt and mutant proteins showed similar localization in the endoplasmic reticulum and Golgi compartments and were transported to the SC surface. These results suggest that the N-glycan of PMP22 facilitates, in part, the stability of the PMP22 oligomer; however, the implications of PMP22 oligomerization remain unknown.
Ryan
M C
MC
Department of Neurobiology, Stanford University School of Medicine, Stanford, California 94305-5125, USA.
Notterpek
L
L
Tobler
A R
AR
Liu
N
N
Shooter
E M
EM
D019943:Amino Acid Substitution; D000818:Animals; D001704:Biopolymers / Q000378:metabolism; D015153:Blotting, Western; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D015870:Gene Expression; D006031:Glycosylation; D006207:Half-Life; D006388:Hemagglutinins / Q000235:genetics; D007150:Immunohistochemistry; D007424:Intracellular Fluid / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009185:Myelin Proteins / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D011233:Precipitin Tests; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012583:Schwann Cells / Q000166:cytology / Q000378:metabolism; D014162:Transfection
D001704:Biopolymers; D006388:Hemagglutinins; D009185:Myelin Proteins; C489546:Pmp22 protein, rat; D011134:Polysaccharides; D016271:Proto-Oncogene Proteins c-myc; D011993:Recombinant Fusion Proteins