A quantitative assay to measure the interaction between immunogenic peptides and purified class I major histocompatibility complex molecules.

TY - JOUR

T1 - A quantitative assay to measure the interaction between immunogenic peptides and purified class I major histocompatibility complex molecules.

AU - Olsen, A C

AU - Pedersen, L O

AU - Hansen, A S

AU - Nissen, Mogens Holst

AU - Olsen, M

AU - Hansen, P R

AU - Holm, A

AU - Buus, S

N1 - Keywords: Amino Acid Sequence; Animals; Antigens, Viral; Binding, Competitive; Cell Line; Histocompatibility Antigens Class I; Mice; Mice, Inbred AKR; Mice, Inbred BALB C; Mice, Inbred C57BL; Molecular Sequence Data; Peptides; Protein Binding; Temperature; Time Factors; beta 2-Microglobulin

PY - 1994

Y1 - 1994

N2 - A direct and sensitive biochemical assay to measure the interaction in solution between peptides and affinity-purified major histocompatibility complex (MHC) class I molecules has been generated. Specific binding reflecting the known class I restriction of cytotoxic T cell responses was obtained. Adding an excess of beta 2-microglobulin (beta 2m) significantly increased the rate of peptide association, but it did not affect the rate of dissociation. Binding was complicated by a rapid and apparently irreversible loss of functional MHC class I at 37 degrees C which might limit the life span of empty MHC class I thereby preventing the inadvertent exchange of peptides at the target cell surface. All class I molecules tested bound peptides of the canonical octa- to nona-meric length. However, one class I molecule, Kk, also bound peptides, which were much longer suggesting that the preference of class I molecules for short epitopes is not absolute and may be caused by factors other than the peptide-MHC class I binding event itself.

AB - A direct and sensitive biochemical assay to measure the interaction in solution between peptides and affinity-purified major histocompatibility complex (MHC) class I molecules has been generated. Specific binding reflecting the known class I restriction of cytotoxic T cell responses was obtained. Adding an excess of beta 2-microglobulin (beta 2m) significantly increased the rate of peptide association, but it did not affect the rate of dissociation. Binding was complicated by a rapid and apparently irreversible loss of functional MHC class I at 37 degrees C which might limit the life span of empty MHC class I thereby preventing the inadvertent exchange of peptides at the target cell surface. All class I molecules tested bound peptides of the canonical octa- to nona-meric length. However, one class I molecule, Kk, also bound peptides, which were much longer suggesting that the preference of class I molecules for short epitopes is not absolute and may be caused by factors other than the peptide-MHC class I binding event itself.

M3 - Journal article

C2 - 8299688

SN - 0014-2980

VL - 24

SP - 385

EP - 392

JO - European Journal of Immunology

JF - European Journal of Immunology

IS - 2

ER -