An essential function of tapasin in quality control of HLA-G molecules.
Tapasin plays an important role in the quality control of major histocompatibility complex (MHC) class I assembly, but its precise function in this process remains controversial. Whether tapasin participates in the assembly of HLA-G has not been studied. HLA-G, an MHC class Ib molecule that binds a more restricted set of peptides than class Ia molecules, is a particularly interesting molecule, because during assembly, it recycles between the endoplasmic reticulum (ER) and the cis-Golgi until it is loaded with a high affinity peptide. We have taken advantage of this unusual trafficking property of HLA-G and its requirement for high affinity peptides to demonstrate that a critical function of tapasin is to transform class I molecules into a high affinity, peptide-receptive form. In the absence of tapasin, HLA-G molecules cannot bind high affinity peptides, and an abundant supply of peptides cannot overcome the tapasin requirement for high affinity peptide loading. The addition of tapasin renders HLA-G molecules capable of loading high affinity peptides and of transporting to the surface, suggesting that tapasin is a prerequisite for the binding of high-affinity ligands. Interestingly, the "tapasin-dependent" HLA-G molecules are not empty in the absence of tapasin but are in fact associated with suboptimal peptides and continue to recycle between the ER and the cis-Golgi. Together with the finding that empty HLA-G heterodimers are strictly retained in the ER and degraded, our data suggest that MHC class I molecules bind any available peptides to avoid ER-mediated degradation and that the peptides are in turn replaced by higher affinity peptides with the aid of tapasin.
Pubmed ID: 12582157 RIS Download
3T3 Cells | Animals | Antigens | Antiporters | Blotting, Western | Cell Membrane | Dimerization | Dose-Response Relationship, Drug | Endoplasmic Reticulum | Flow Cytometry | Golgi Apparatus | HLA Antigens | HLA-G Antigens | Histocompatibility Antigens Class I | Humans | Immunoglobulins | Kinetics | Ligands | Membrane Transport Proteins | Mice | Microscopy, Fluorescence | Microsomes | Peptides | Precipitin Tests | Protein Conformation | Temperature