title: Biochemical specificity of H-2M3a. Stereospecificity and space-filling requirements at position 1 maintain N-formyl peptide binding.
abstract: The maternally transmitted Ag is a cell surface product of three gene products: 1) H-2M3a (formerly Hmta), a class I MHC heavy chain; 2) beta 2-microglobulin; and 3) maternally transmitted factor (Mtf), the N-terminus of the mitochondrially encoded ND1 subunit of the reduced form of nicotinamide-adenine dinucleotide dehydrogenase. This class I molecule has been shown to be an N-formyl peptide receptor. Although the N-formyl moiety is necessary for binding to M3a, it is not sufficient. We proposed that the R group of the amino acid in position 1 plays a pivotal role in peptide binding to M3a. To test this hypothesis, analogues differing in size and stereospecificity of the R group were synthesized. Substitutions with other hydrophobic amino acids such as N-formyl phenylalanine and N-formyl valine had no significant effect on the ability of these Mtf alpha analogues to sensitize target cells (M3a, Mtf beta) to M3a, Mtf alpha-specific CTL. In contrast, the nonsubstituted, N-formylated, and N-acetylated glycyl analogues of Mtf beta bound equivalently to M3a in a peptide competition assay. Moreover, the alanine analogue bound in an N-formyl-dependent manner. To determine the limitations of the putative N-formyl pocket, peptide analogues were constructed incorporating D-isomer amino acids. When formylated D-alanine or D-methionine replaced the native methionine, these peptide derivatives did not show significant binding to M3a. Therefore, the presence of a space-filling R group (greater than hydrogen) is necessary for an antigenic peptide to bind M3a in an N-formyl-dependent manner. Additionally, the ability of M3a to discriminate between the optical forms of methionine and alanine demonstrates that this N-formyl pocket is stereospecific in its ability to bind peptide. Thus, we have defined three requirements for peptide binding to M3a: an N-formyl moiety at the amino terminus of the peptide, a space-filling R group at position 1 to maintain this N-formyl specificity, and the correct stereoisomer of the first amino acid.
authors.affiliation: Department of Microbiology and Immunology, Baylor College of Medicine, Houston, TX 77030.
authors.forename: J M
authors.initials: JM
authors.lastname: Vyas
authors.forename: S M
authors.initials: SM
authors.lastname: Shawar
authors.forename: J R
authors.initials: JR
authors.lastname: Rodgers
authors.forename: R G
authors.initials: RG
authors.lastname: Cook
authors.forename: R R
authors.initials: RR
authors.lastname: Rich
mesh_terms: D000409:Alanine / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D005998:Glycine / Q000737:chemistry; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009038:Motion; D009239:N-Formylmethionine / Q000378:metabolism*; D010455:Peptides / Q000276:immunology*; D011485:Protein Binding; D013237:Stereoisomerism; D013329:Structure-Activity Relationship
chemical_list: C078177:H2-M3 antigen; D015395:Histocompatibility Antigens Class I; D010455:Peptides; D009239:N-Formylmethionine; D000409:Alanine; D005998:Glycine