Effects of β₂–microglobulin mutations on the alpha-1 helical region of H2-L^d

Abstract

 Beta-2 microglobulin (β₂m)has been shown to have an effect on the structural and functional constraints that facilitate proper class I antigen presentation. To date, no evidence has pinpointed the β₂m-specific amino acids that play an integral role in affecting structure in and around the peptide binding region of class I. To delineate β₂m amino acid positions that affect the alpha-1 helical region, we generated a series of mutant β₂m proteins bearing precise amino acid substitutions. The amino acid positions chosen were based upon previous results which demonstrated that human β₂m association with H2-L^d altered the structure of the alpha-1/alpha-2 super-domain. β₂m mutant proteins were used in β₂m exchange assays with cells expressing H2-L^d. Following exchange, cells were assayed to determine whether mutant β₂m association resulted in structural alteration of class I extracellular domains. The alteration in H2-L^d structure was evidenced by an increase in the binding of an antibody (34-1-2), specific for the alpha-1 helical region of H2-L^d. Results demonstrated that amino acid substitutions in β₂m positions 33 and 53 led to a dramatic increase in the reactivity of the alpha-1 domain-specific antibody 34-1-2. Identifying β₂m amino acid positions that influence the structure of the peptide binding region may allow for a better understanding of cellular immune responses that center upon class I/β₂m expression.