title: Androgen receptor ligand-binding domain interaction and nuclear receptor specificity of FXXLF and LXXLL motifs as determined by L/F swapping.
abstract: The androgen receptor (AR) ligand-binding domain (LBD) binds FXXLF motifs, present in the AR N-terminal domain and AR-specific cofactors, and some LXXLL motifs of nuclear receptor coactivators. We demonstrated that in the context of the AR FXXLF motif many different amino acid residues at positions +2 and +3 are compatible with strong AR LBD interaction, although a preference for E at +2 and K or R at +3 was found. Pairwise systematic analysis of F/L swaps at +1 and +5 in FXXLF and LXXLL motifs showed: 1) F to L substitutions in natural FXXLF motifs abolished AR LBD interaction; 2) binding of interacting LXXLL motifs was unchanged or increased upon L to F substitutions; 3) certain noninteracting LXXLL motifs became strongly AR-interacting FXXLF motifs; whereas 4) other nonbinders remained unaffected by L to F substitutions. All FXXLF motifs, but not the corresponding LXXLL motifs, displayed a strong preference for AR LBD. Progesterone receptor LBD interacted with some FXXLF motifs, albeit always less efficiently than corresponding LXXLL motifs. AR LBD interaction of most FXXLF and LXXLL peptides depended on classical charge clamp residue K720, whereas E897 was less important. Other charged residues lining the AR coactivator-binding groove, K717 and R726, modulated optimal peptide binding. Interestingly, these four charged residues affected binding of individual peptides independent of an F or L at +1 and +5 in swap experiments. In conclusion, F residues determine strong and selective peptide interactions with AR. Sequences flanking the core motif determine the specific mode of FXXLF and LXXLL interactions.
authors.affiliation: Department of Pathology, Josephine Nefkens Institute, Erasmus MC, Rotterdam, The Netherlands. h.dubbink@erasmusmc.nl
authors.forename: Hendrikus J
authors.initials: HJ
authors.lastname: Dubbink
authors.forename: Remko
authors.initials: R
authors.lastname: Hersmus
authors.forename: Ashley C W
authors.initials: AC
authors.lastname: Pike
authors.forename: Michel
authors.initials: M
authors.lastname: Molier
authors.forename: Albert O
authors.initials: AO
authors.lastname: Brinkmann
authors.forename: Guido
authors.initials: G
authors.lastname: Jenster
authors.forename: Jan
authors.initials: J
authors.lastname: Trapman
mesh_terms: D020816:Amino Acid Motifs / Q000502:physiology; D001665:Binding Sites; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D019281:Dimerization; D051548:Histone Acetyltransferases; D006801:Humans; D007930:Leucine / Q000378:metabolism*; D016350:Leucine Zippers / Q000502:physiology*; D008024:Ligands; D008958:Models, Molecular; D056920:Nuclear Receptor Coactivator 1; D051958:Nuclear Receptor Coactivator 2 / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D010649:Phenylalanine / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D011522:Protons; D011944:Receptors, Androgen / Q000737:chemistry* / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D020798:Two-Hybrid System Techniques
chemical_list: D008024:Ligands; C498001:NCOA2 protein, human; D051958:Nuclear Receptor Coactivator 2; D010455:Peptides; D011522:Protons; D011944:Receptors, Androgen; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors; D010649:Phenylalanine; D051548:Histone Acetyltransferases; C538859:NCOA1 protein, human; D056920:Nuclear Receptor Coactivator 1; D007930:Leucine