Ribophorin II, also known as dolichyl-diphosphooligosaccharide–protein glycosyltransferase subunit 2, is a crucial component of the N-oligosaccharyl transferase complex. This complex is responsible for the attachment of high mannose oligosaccharides to asparagine residues in nascent polypeptide chains, specifically within the Asn-X-Ser/Thr consensus motif .
Ribophorin II is a non-catalytic subunit of the oligosaccharyltransferase complex. It plays a significant role in the glycosylation process, which is essential for proper protein folding and stability. The human recombinant form of Ribophorin II is produced in Escherichia coli and is a single, non-glycosylated polypeptide chain containing 539 amino acids, with a molecular mass of approximately 59.2 kilodaltons .
The recombinant Ribophorin II is fused to a 21 amino acid His-tag at the N-terminus, which facilitates its purification through proprietary chromatographic techniques. The protein is typically formulated in a solution containing 20 millimolar Tris-HCl buffer (pH 8.0), 10% glycerol, and 0.1 molar sodium chloride .
For optimal stability, Ribophorin II should be stored at 4°C if it will be used within 2-4 weeks. For longer storage periods, it is recommended to keep the protein frozen at -20°C, with the addition of a carrier protein such as human serum albumin or bovine serum albumin to prevent degradation. It is important to avoid multiple freeze-thaw cycles to maintain the protein’s integrity .
Ribophorin II is highly conserved across various species, indicating its essential role in cellular processes. It is involved in the glycosylation of proteins, a critical post-translational modification that affects protein folding, stability, and function. The protein’s importance is underscored by its association with several diseases, including progressive encephalopathy with brain atrophy and spasticity, and hereditary sensory and autonomic neuropathy type VII .
Recombinant Ribophorin II is widely used in laboratory research to study protein glycosylation and its implications in various biological processes and diseases. Its high purity and stability make it a valuable tool for researchers investigating the mechanisms of protein folding and the role of glycosylation in health and disease .