The role of protein-protein and protein-membrane Interactions on P450 function

Emily E. Scott, C. Roland Wolf, Michal Otyepka, Sara C. Humphreys, James R. Reed, Colin J. Henderson, Lesley A. McLaughlin, Marketa Paloncyova, Veronika Navratilova, Karel Berka, Pavel Anzenbacher, Upendra P. Dahal, Carlo Barnaba, James A. Brozik, Jeffrey P. Jones, Fernando Estrada, Jennifer S. Laurence, Ji Won Park, Wayne L. Backes (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    19 Citations (Scopus)

    Abstract

    This symposium summary, sponsored by the ASPET, was held at Experimental Biology 2015 on March 29th in Boston MA. The symposium focused on (1) the interactions of P450s with their redox partners, and (2) the role of the lipid membrane in their orientation and stabilization. Two presentations discussed the interactions of P450s with NADPH-cytochrome P450 reductase (CPR) and cytochrome b5. First, solution NMR was used to compare the protein interactions that facilitated either the hydroxylase or lyase activities of CYP17A1. The lyase interaction was stimulated by the presence of b5 and 17α-hydroxypregnenolone, whereas the hydroxylase reaction was predominant in the absence of b5. The role of b5 was also shown in vivo by selective hepatic knockout of b5 from mice expressing CYP3A4 and CYP2D6 - the lack of b5 caused a decrease in the clearance of several substrates. The role of the membrane on P450 orientation was examined using computational methodologies, showing that the proximal region of the P450 molecule faced the aqueous phase. The distal region, containing the substrate-access channel, was associated with the membrane. The interaction of CPR with the membrane was also described, showing the ability of CPR to "helicopter" above the membrane. Finally, the endoplasmic reticulum (ER) was shown to be heterogeneous, having ordered membrane regions containing cholesterol and more disordered regions. Interestingly, two closely-related CYPs, CYP1A1 and CYP1A2 resided in different regions of the ER. The structural characteristics of their localization were examined. These studies emphasize the importance of P450 protein organization to their function.

    Original languageEnglish
    Pages (from-to)576-590
    Number of pages15
    JournalDrug Metabolism and Disposition
    Volume44
    Issue number4
    Early online date5 Feb 2016
    DOIs
    Publication statusPublished - Apr 2016

    Fingerprint

    Membrane Proteins
    NADPH-Ferrihemoprotein Reductase
    Membranes
    Lyases
    Mixed Function Oxygenases
    Proteins
    Endoplasmic Reticulum
    17-alpha-Hydroxypregnenolone
    Cytochromes b5
    Cytochrome P-450 CYP3A
    Cytochrome P-450 CYP1A2
    Cytochrome P-450 CYP2D6
    Cytochrome P-450 CYP1A1
    Aircraft
    Membrane Lipids
    Knockout Mice
    Oxidation-Reduction
    Cholesterol
    Liver

    Keywords

    • animal models
    • cytochrome b5
    • enzyme mechanism
    • enzyme structure
    • membrane-protein interactions
    • NADPH cytochrome P450 reductase
    • pharmacokinetics
    • protein-protein interactions
    • reductases

    Cite this

    Scott, Emily E. ; Wolf, C. Roland ; Otyepka, Michal ; Humphreys, Sara C. ; Reed, James R. ; Henderson, Colin J. ; McLaughlin, Lesley A. ; Paloncyova, Marketa ; Navratilova, Veronika ; Berka, Karel ; Anzenbacher, Pavel ; Dahal, Upendra P. ; Barnaba, Carlo ; Brozik, James A. ; Jones, Jeffrey P. ; Estrada, Fernando ; Laurence, Jennifer S. ; Park, Ji Won ; Backes, Wayne L. / The role of protein-protein and protein-membrane Interactions on P450 function. In: Drug Metabolism and Disposition. 2016 ; Vol. 44, No. 4. pp. 576-590.
    @article{cc65f1f5cbfa47c7b35fbf89b50dbbec,
    title = "The role of protein-protein and protein-membrane Interactions on P450 function",
    abstract = "This symposium summary, sponsored by the ASPET, was held at Experimental Biology 2015 on March 29th in Boston MA. The symposium focused on (1) the interactions of P450s with their redox partners, and (2) the role of the lipid membrane in their orientation and stabilization. Two presentations discussed the interactions of P450s with NADPH-cytochrome P450 reductase (CPR) and cytochrome b5. First, solution NMR was used to compare the protein interactions that facilitated either the hydroxylase or lyase activities of CYP17A1. The lyase interaction was stimulated by the presence of b5 and 17α-hydroxypregnenolone, whereas the hydroxylase reaction was predominant in the absence of b5. The role of b5 was also shown in vivo by selective hepatic knockout of b5 from mice expressing CYP3A4 and CYP2D6 - the lack of b5 caused a decrease in the clearance of several substrates. The role of the membrane on P450 orientation was examined using computational methodologies, showing that the proximal region of the P450 molecule faced the aqueous phase. The distal region, containing the substrate-access channel, was associated with the membrane. The interaction of CPR with the membrane was also described, showing the ability of CPR to {"}helicopter{"} above the membrane. Finally, the endoplasmic reticulum (ER) was shown to be heterogeneous, having ordered membrane regions containing cholesterol and more disordered regions. Interestingly, two closely-related CYPs, CYP1A1 and CYP1A2 resided in different regions of the ER. The structural characteristics of their localization were examined. These studies emphasize the importance of P450 protein organization to their function.",
    keywords = "animal models , cytochrome b5, enzyme mechanism, enzyme structure, membrane-protein interactions, NADPH cytochrome P450 reductase, pharmacokinetics, protein-protein interactions, reductases",
    author = "Scott, {Emily E.} and Wolf, {C. Roland} and Michal Otyepka and Humphreys, {Sara C.} and Reed, {James R.} and Henderson, {Colin J.} and McLaughlin, {Lesley A.} and Marketa Paloncyova and Veronika Navratilova and Karel Berka and Pavel Anzenbacher and Dahal, {Upendra P.} and Carlo Barnaba and Brozik, {James A.} and Jones, {Jeffrey P.} and Fernando Estrada and Laurence, {Jennifer S.} and Park, {Ji Won} and Backes, {Wayne L.}",
    note = "The American Society for Pharmacology and Experimental Therapeutics.",
    year = "2016",
    month = "4",
    doi = "10.1124/dmd.115.068569",
    language = "English",
    volume = "44",
    pages = "576--590",
    journal = "Drug Metabolism and Disposition",
    issn = "0090-9556",
    publisher = "American Society for Pharmacology and Experimental Therapeutics",
    number = "4",

    }

    Scott, EE, Wolf, CR, Otyepka, M, Humphreys, SC, Reed, JR, Henderson, CJ, McLaughlin, LA, Paloncyova, M, Navratilova, V, Berka, K, Anzenbacher, P, Dahal, UP, Barnaba, C, Brozik, JA, Jones, JP, Estrada, F, Laurence, JS, Park, JW & Backes, WL 2016, 'The role of protein-protein and protein-membrane Interactions on P450 function', Drug Metabolism and Disposition, vol. 44, no. 4, pp. 576-590. https://doi.org/10.1124/dmd.115.068569

    The role of protein-protein and protein-membrane Interactions on P450 function. / Scott, Emily E.; Wolf, C. Roland; Otyepka, Michal; Humphreys, Sara C.; Reed, James R.; Henderson, Colin J.; McLaughlin, Lesley A.; Paloncyova, Marketa; Navratilova, Veronika; Berka, Karel; Anzenbacher, Pavel; Dahal, Upendra P.; Barnaba, Carlo; Brozik, James A.; Jones, Jeffrey P.; Estrada, Fernando; Laurence, Jennifer S.; Park, Ji Won; Backes, Wayne L. (Lead / Corresponding author).

    In: Drug Metabolism and Disposition, Vol. 44, No. 4, 04.2016, p. 576-590.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - The role of protein-protein and protein-membrane Interactions on P450 function

    AU - Scott, Emily E.

    AU - Wolf, C. Roland

    AU - Otyepka, Michal

    AU - Humphreys, Sara C.

    AU - Reed, James R.

    AU - Henderson, Colin J.

    AU - McLaughlin, Lesley A.

    AU - Paloncyova, Marketa

    AU - Navratilova, Veronika

    AU - Berka, Karel

    AU - Anzenbacher, Pavel

    AU - Dahal, Upendra P.

    AU - Barnaba, Carlo

    AU - Brozik, James A.

    AU - Jones, Jeffrey P.

    AU - Estrada, Fernando

    AU - Laurence, Jennifer S.

    AU - Park, Ji Won

    AU - Backes, Wayne L.

    N1 - The American Society for Pharmacology and Experimental Therapeutics.

    PY - 2016/4

    Y1 - 2016/4

    N2 - This symposium summary, sponsored by the ASPET, was held at Experimental Biology 2015 on March 29th in Boston MA. The symposium focused on (1) the interactions of P450s with their redox partners, and (2) the role of the lipid membrane in their orientation and stabilization. Two presentations discussed the interactions of P450s with NADPH-cytochrome P450 reductase (CPR) and cytochrome b5. First, solution NMR was used to compare the protein interactions that facilitated either the hydroxylase or lyase activities of CYP17A1. The lyase interaction was stimulated by the presence of b5 and 17α-hydroxypregnenolone, whereas the hydroxylase reaction was predominant in the absence of b5. The role of b5 was also shown in vivo by selective hepatic knockout of b5 from mice expressing CYP3A4 and CYP2D6 - the lack of b5 caused a decrease in the clearance of several substrates. The role of the membrane on P450 orientation was examined using computational methodologies, showing that the proximal region of the P450 molecule faced the aqueous phase. The distal region, containing the substrate-access channel, was associated with the membrane. The interaction of CPR with the membrane was also described, showing the ability of CPR to "helicopter" above the membrane. Finally, the endoplasmic reticulum (ER) was shown to be heterogeneous, having ordered membrane regions containing cholesterol and more disordered regions. Interestingly, two closely-related CYPs, CYP1A1 and CYP1A2 resided in different regions of the ER. The structural characteristics of their localization were examined. These studies emphasize the importance of P450 protein organization to their function.

    AB - This symposium summary, sponsored by the ASPET, was held at Experimental Biology 2015 on March 29th in Boston MA. The symposium focused on (1) the interactions of P450s with their redox partners, and (2) the role of the lipid membrane in their orientation and stabilization. Two presentations discussed the interactions of P450s with NADPH-cytochrome P450 reductase (CPR) and cytochrome b5. First, solution NMR was used to compare the protein interactions that facilitated either the hydroxylase or lyase activities of CYP17A1. The lyase interaction was stimulated by the presence of b5 and 17α-hydroxypregnenolone, whereas the hydroxylase reaction was predominant in the absence of b5. The role of b5 was also shown in vivo by selective hepatic knockout of b5 from mice expressing CYP3A4 and CYP2D6 - the lack of b5 caused a decrease in the clearance of several substrates. The role of the membrane on P450 orientation was examined using computational methodologies, showing that the proximal region of the P450 molecule faced the aqueous phase. The distal region, containing the substrate-access channel, was associated with the membrane. The interaction of CPR with the membrane was also described, showing the ability of CPR to "helicopter" above the membrane. Finally, the endoplasmic reticulum (ER) was shown to be heterogeneous, having ordered membrane regions containing cholesterol and more disordered regions. Interestingly, two closely-related CYPs, CYP1A1 and CYP1A2 resided in different regions of the ER. The structural characteristics of their localization were examined. These studies emphasize the importance of P450 protein organization to their function.

    KW - animal models

    KW - cytochrome b5

    KW - enzyme mechanism

    KW - enzyme structure

    KW - membrane-protein interactions

    KW - NADPH cytochrome P450 reductase

    KW - pharmacokinetics

    KW - protein-protein interactions

    KW - reductases

    U2 - 10.1124/dmd.115.068569

    DO - 10.1124/dmd.115.068569

    M3 - Article

    C2 - 26851242

    VL - 44

    SP - 576

    EP - 590

    JO - Drug Metabolism and Disposition

    JF - Drug Metabolism and Disposition

    SN - 0090-9556

    IS - 4

    ER -