Differences in conformational stability of the two alpha domains of the disease-associated and non-disease-associated subtypes of HLA-B27.
The MHC Class I molecule, HLA-B27, is strongly linked with development of the inflammatory arthritic disease, ankylosing spondylitis (AS); whereas the B*2705 subtype shows strong association, B*2709 is not associated with disease, even though the two subtypes differ in only a single residue at position 116. Currently, attention is focused on the misfolding propensities of these two subtypes, including studies of disulfide-linked dimers and non-covalently formed high molecular weight (HMW) aggregates. Using mutants retaining only a single cysteine at positions C67 or C164, and using a cysteine-reactive, environment-sensitive, fluorescence probe (acrylodan), we find that within the same overall population of identical single-cysteine HLA-B27 molecules, there exist sub-populations which (a) possess free cysteines which react with acrylodan, (b) form disulfide-linked dimers, and (c) form HMW aggregates. Further, using acrylodan fluorescence, we find (d) that the α1 and α2 domains unfold independently of each other in HMW aggregates, (e) that these two domains of B*2709 are less stable to chemical and thermal denaturation than the corresponding domains of B*2705, suggesting easier clearance of misfolded molecules in the former, and (f) C67 is much more exposed in B*2705 than in B*2709, which could potentially explain how B*2705 more easily forms C67-mediated disulfide-bonded dimers.
Department of Immunopathology, Postgraduate Institute of Medical Education and Research (PGIMER), Sector-12, Chandigarh - 160012, India.
Manish Kumar
MK
Rana
Department of Immunopathology, Postgraduate Institute of Medical Education and Research (PGIMER), Sector-12, Chandigarh - 160012, India. Electronic address: guptasarma.manni@pgimer.edu.in.
Manni
M
Luthra-Guptasarma
D015796:HLA-B27 Antigen / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008970:Molecular Weight; D020125:Mutation, Missense; D011489:Protein Denaturation; D000072417:Protein Domains; D055550:Protein Stability; D013167:Spondylitis, Ankylosing / Q000235:genetics; D014508:Urea / Q000737:chemistry
D015796:HLA-B27 Antigen; D014508:Urea