5FA6

wild type human CYPOR

Summary for 5FA6

• Entry DOI: 10.2210/pdb5fa6/pdb
• Related: 3QE2
• Descriptor: NADPH--cytochrome P450 reductase, FLAVIN MONONUCLEOTIDE, FLAVIN-ADENINE DINUCLEOTIDE, ... (5 entities in total)
• Functional Keywords: cypor, cytochrome p450 reductase, flavoprotein, abs-like phenotype, oxidoreductase
• Biological source: Homo sapiens (Human)
• Cellular location: Endoplasmic reticulum membrane ; Single-pass membrane protein ; Cytoplasmic side : P16435
• Total number of polymer chains: 2
• Total formula weight: 144142.35

Authors

Xia, C.,Marohnic, C.,Panda, S.,Masters, B.S.,Kim, J.J.K. (deposition date: 2015-12-11, release date: 2016-08-17, Last modification date: 2024-03-06)

Primary citation

McCammon, K.M.,Panda, S.P.,Xia, C.,Kim, J.J.,Moutinho, D.,Kranendonk, M.,Auchus, R.J.,Lafer, E.M.,Ghosh, D.,Martasek, P.,Kar, R.,Masters, B.S.,Roman, L.J.
Instability of the Human Cytochrome P450 Reductase A287P Variant Is the Major Contributor to Its Antley-Bixler Syndrome-like Phenotype.
J.Biol.Chem., 291:20487-20502, 2016

PubMed Abstract: Human NADPH-cytochrome P450 oxidoreductase (POR) gene mutations are associated with severe skeletal deformities and disordered steroidogenesis. The human POR mutation A287P presents with disordered sexual development and skeletal malformations. Difficult recombinant expression and purification of this POR mutant suggested that the protein was less stable than WT. The activities of cytochrome P450 17A1, 19A1, and 21A2, critical in steroidogenesis, were similar using our purified, full-length, unmodified A287P or WT POR, as were those of several xenobiotic-metabolizing cytochromes P450, indicating that the A287P protein is functionally competent in vitro, despite its functionally deficient phenotypic behavior in vivo Differential scanning calorimetry and limited trypsinolysis studies revealed a relatively unstable A287P compared with WT protein, leading to the hypothesis that the syndrome observed in vivo results from altered POR protein stability. The crystal structures of the soluble domains of WT and A287P reveal only subtle differences between them, but these differences are consistent with the differential scanning calorimetry results as well as the differential susceptibility of A287P and WT observed with trypsinolysis. The relative in vivo stabilities of WT and A287P proteins were also examined in an osteoblast cell line by treatment with cycloheximide, a protein synthesis inhibitor, showing that the level of A287P protein post-inhibition is lower than WT and suggesting that A287P may be degraded at a higher rate. Current studies demonstrate that, unlike previously described mutations, A287P causes POR deficiency disorder due to conformational instability leading to proteolytic susceptibility in vivo, rather than through an inherent flavin-binding defect.

PubMed: 27496950
DOI: 10.1074/jbc.M116.716019

Experimental method

X-RAY DIFFRACTION (2.3 Å)