TY - JOUR
T1 - The first structure of a protein from the SOUL/HBP family of heme-binding proteins: Murine p22HBP
AU - Dias, Jorge da Silva
AU - Macedo, Maria dos Anjos López de
AU - Ferreira, Glória C.
AU - Peterson, Francis C.
AU - Volkman, Brian F.
AU - Goodfellow, Brian J.
PY - 2006/10/20
Y1 - 2006/10/20
N2 - Murine p22HBP, a 22-kDa monomer originally identified as a cytosolic heme-binding protein ubiquitously expressed in various tissues, has 27% sequence identity to murine SOUL, a heme-binding hexamer specifically expressed in the retina. In contrast to murine SOUL, which binds one heme per subunit via coordination of the Fe(III)-heme to a histidine, murine p22HBP binds one heme molecule per subunit with no specific axial ligand coordination of the Fe(III)-heme. Using intrinsic protein fluorescence quenching, the values for the dissociation constants of p22HBP for hemin and protoporphyrin-IX were determined to be in the low nanomolar range. The three-dimensional structure of murine p22HBP, the first for a protein from the SOUL/HBP family, was determined by NMR methods to consist of a 9-stranded distorted β-barrel flanked by two long α-helices. Although homologous domains have been found in three bacterial proteins, two of which are transcription factors, the fold determined for p22HBP corresponds to a novel α plus β fold in a eukaryotic protein. Chemical shift mapping localized the tetrapyrrole binding site to a hydrophobic cleft formed by residues from helix αA and an extended loop. In an attempt to assess the structural basis for tetrapyrrole binding in the SOUL/HBP family, models for the p22HBP-protoporphyrin-IX complex and the SOUL protein were generated by manual docking and automated methods.
AB - Murine p22HBP, a 22-kDa monomer originally identified as a cytosolic heme-binding protein ubiquitously expressed in various tissues, has 27% sequence identity to murine SOUL, a heme-binding hexamer specifically expressed in the retina. In contrast to murine SOUL, which binds one heme per subunit via coordination of the Fe(III)-heme to a histidine, murine p22HBP binds one heme molecule per subunit with no specific axial ligand coordination of the Fe(III)-heme. Using intrinsic protein fluorescence quenching, the values for the dissociation constants of p22HBP for hemin and protoporphyrin-IX were determined to be in the low nanomolar range. The three-dimensional structure of murine p22HBP, the first for a protein from the SOUL/HBP family, was determined by NMR methods to consist of a 9-stranded distorted β-barrel flanked by two long α-helices. Although homologous domains have been found in three bacterial proteins, two of which are transcription factors, the fold determined for p22HBP corresponds to a novel α plus β fold in a eukaryotic protein. Chemical shift mapping localized the tetrapyrrole binding site to a hydrophobic cleft formed by residues from helix αA and an extended loop. In an attempt to assess the structural basis for tetrapyrrole binding in the SOUL/HBP family, models for the p22HBP-protoporphyrin-IX complex and the SOUL protein were generated by manual docking and automated methods.
KW - Protein fluorescence quenching
KW - Retina
KW - Ligands
KW - Protoporphyrin
UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-33846009114&origin=inward&txGid=05d117a793aa01d433e544a4b97a1134#
U2 - 10.1074/jbc.M605988200
DO - 10.1074/jbc.M605988200
M3 - Article
SN - 0021-9258
VL - 281
SP - 31553
EP - 31561
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - NA
ER -