Cytoplasmic glycosylation of protein-hydroxyproline and its relationship to other glycosylation pathways.
The Skp1 protein, best known as a subunit of E3(SCF)-ubiquitin ligases, is subject to complex glycosylation in the cytoplasm of the cellular slime mold Dictyostelium. Pro143 of this protein is sequentially modified by a prolyl hydroxylase and five soluble glycosyltransferases (GT), to yield the structure Galalpha1,Galalpha1,3Fucalpha1,2Galbeta1,3GlcNAcalpha1-HyPro143. These enzymes are unusual in that they are expressed in the cytoplasmic compartment of the cell, rather than the secretory pathway where complex glycosylation of proteins usually occurs. The first enzyme in the pathway appears to be related to the soluble animal prolyl 4-hydroxylases (P4H), which modify the transcriptional factor subunit HIF-1alpha in the cytoplasm, and more distantly to the P4Hs that modify collagen and other proteins in the rER, based on biochemical and informatics analyses. The soluble alphaGlcNAc-transferase acting on Skp1 has been cloned and is distantly related to the mucin-type polypeptide N-acetyl-alpha-galactosaminyltransferase in the Golgi of animals. Its characterization has led to the discovery of a family of related polypeptide N-acetyl-alpha-glucosaminyltransferases in the Golgi of selected lower eukaryotes. The Skp1 GlcNAc is extended by a bifunctional diglycosyltransferase that sequentially and apparently processively adds beta1,3Gal and alpha1,2Fuc. Though this structure is also formed in the animal secretory pathway, the GTs involved are dissimilar. Conceptual translation of available genomes suggests the existence of this kind of complex cytoplasmic glycosylation in other eukaryotic microorganisms, including diatoms, oomycetes, and possibly Chlamydomonas and Toxoplasma, and an evolutionary precursor of this pathway may also occur in prokaryotes.
West
Christopher M
CM
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA. Christopher-west@ouhsc.edu
Van Der Wel
Hanke
H
Sassi
Slim
S
Gaucher
Eric A
EA
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003593:Cytoplasm / Q000201:enzymology / Q000378:metabolism*; D004023:Dictyostelium / Q000235:genetics / Q000378:metabolism; D005700:Galactosyltransferases / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D011393:Procollagen-Proline Dioxygenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D044786:S-Phase Kinase-Associated Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D014157:Transcription Factors / Q000378:metabolism
C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D011134:Polysaccharides; D044786:S-Phase Kinase-Associated Proteins; D014157:Transcription Factors; D011393:Procollagen-Proline Dioxygenase; D005700:Galactosyltransferases; D017351:N-Acetylglucosaminyltransferases; C403138:UDP-N-acetylglucosamine - SKP1-hydroxyproline N-acetylglucosaminyltransferase; C120745:UDP-galactose N-acetylglucosaminyl-1-3-N-acetylgalactosamine beta-1,3-galactosyltransferase