Structural features and dynamic investigations of the membrane-bound cytochrome P450 17A1

Cui, Ying; Xue, Qiao; Zheng, Qing‐Chuan; Zhang, Jilong; Kong, Chui‐Peng; Fan, Junwen; Zhang, Hongxing

doi:10.1016/j.bbamem.2015.05.017

Cited by 18 publications

(27 citation statements)

References 53 publications

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“…To identify protein tunnels, studies of membrane-bound P450s usually use probe cutoffs around 1.2 Å 33,36,73 , which is actually less than the radius of a water molecule (1.4 Å). However, typical organic substrates of P450 family 1 enzymes are aromatic hydrocarbons and their derivatives, with their minimal radius significantly larger than 1.4 Å.…”

Section: Resultsmentioning

confidence: 99%

Lipid molecules can induce an opening of membrane-facing tunnels in cytochrome P450 1A2

Jeřábek

Florián

Martínek

2016

Phys. Chem. Chem. Phys.

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Cytochrome P450 1A2 (P450 1A2, CYP1A2) is a membrane-bound enzyme that oxidizes a broad range of hydrophobic substrates. The structure and dynamics of both the catalytic and trans-membrane (TM) domains of this enzyme in the membrane/water environment were investigated using a multiscale computational approach, including coarse-grained and all-atom molecular dynamics. Starting from the spontaneous self-assembly of the system containing the TM or soluble domain immersed in randomized dilauroyl phosphatidylcholine (DLPC)/water mixture into their respective membrane-bound forms, we reconstituted the membrane-bound structure of the full-length P450 1A2. This structure includes a TM helix that spans the membrane, while being connected to the catalytic domain by a short flexible loop. Furthermore, in this model, the upper part of the TM helix interacts directly with a conserved and highly hydrophobic N-terminal proline-rich segment of the catalytic domain; this segment and the FG loop are immersed in the membrane, whereas the remaining portion of the catalytic domain remain exposed to the aqueous solution. The shallow membrane immersion of the catalytic domain appears to induce a depression in the opposite intact layer of the phospholipids, which may help in stabilizing the position of the TM helix directly beneath the catalytic domain. The partial immersion of the catalytic domain also allows for the enzyme substrates to enter the active site from either aqueous solution or phospholipid environments via several solvent- and membrane-facing tunnels in the full-length P450 1A2. The calculated tunnel dynamics indicated that the opening probability of the membrane-facing tunnels is significantly enhanced when a DLPC molecule spontaneously penetrates into the membrane-facing tunnel 2d. The energetics of the lipid penetration process were assessed by the linear interaction energy (LIE) approximation, and found to be thermodynamically feasible.

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Section: Resultsmentioning

confidence: 99%

Lipid molecules can induce an opening of membrane-facing tunnels in cytochrome P450 1A2

Jeřábek

Florián

Martínek

2016

Phys. Chem. Chem. Phys.

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“…The length of the TM-helix was predicted by the secondary structure prediction servers: PredictProtein (https://www.predictprotein.org/) (21) and PSIPRED (http://bioinf.cs.ucl.ac.uk/psipred/) (22). The TM-helix reported by Cui et al (23) for wtCYP17 extended over residues 1-19. The N-terminal missing regions were modeled in a helical conformation by applying constraints on residues 3-19 and assuming a loop conformation for residues 20-30.…”

Section: Preparation Of All-atom Models Of the Proteinsmentioning

confidence: 99%

“…For mtCYP19, no TM-helix domain was predicted by secondary structure prediction servers. Therefore, the N-terminal residues (36)(37)(38)(39)(40)(41)(42)(43)(44) were modeled as a loop region. To facilitate comparison, the residue numbering of wtCYP19 was retained for mtCYP19.…”

Section: Preparation Of All-atom Models Of the Proteinsmentioning

confidence: 99%

“…They are important for stabilization of the TM-helix in the bilayer and for protein sorting and retention of CYPs in the ER membrane (4). For example, it has been observed that retention of the TM-helix of CYP 2C1 in the ER membrane is dependent on the hydrophobicity of the TM domain (residues [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] and on the hydrophilic residues (21-23 KQS) at the beginning of the linker region (residues [21][22][23][24][25][26][27][28]. Mutation of the linker, the hydrophobic TM domain, or both regions resulted in the expulsion of CYP 2C1 from the ER membrane and affected direct retrieval from the pre-Golgi compartment (5).…”

Section: Introductionmentioning

confidence: 99%

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Influence of Transmembrane Helix Mutations on Cytochrome P450-Membrane Interactions and Function

Mustafa

Nandekar

Camp

et al. 2019

Biophysical Journal

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Human cytochrome P450 (CYP) enzymes play an important role in the metabolism of drugs, steroids, fatty acids, and xenobiotics. Microsomal CYPs are anchored in the endoplasmic reticulum membrane by an N-terminal transmembrane (TM) helix that is connected to the globular catalytic domain by a flexible linker sequence. However, the structural and functional importance of the TM-helix is unclear because it has been shown that CYPs can still associate with the membrane and have enzymatic activity in reconstituted systems after truncation or modification of the N-terminal sequence. Here, we investigated the effect of mutations in the N-terminal TM-helix residues of two human steroidogenic enzymes, CYP 17A1 and CYP 19A1, that are major drug targets for cancer therapy. These mutations were originally introduced to increase the expression of the proteins in Escherichia coli. To investigate the effect of the mutations on protein-membrane interactions and function, we carried out coarse-grained and all-atom molecular dynamics simulations of the CYPs in a phospholipid bilayer. We confirmed the orientations of the globular domain in the membrane observed in the simulations by linear dichroism measurements in a Nanodisc. Whereas the behavior of CYP 19A1 was rather insensitive to truncation of the TM-helix, mutations in the TM-helix of CYP 17A1, especially W2A and E3L, led to a gradual drifting of the TM-helix out of the hydrophobic core of the membrane. This instability of the TM-helix could affect interactions with the allosteric redox partner, cytochrome b5, required for CYP 17A1's lyase activity. Furthermore, the simulations showed that the mutant TM-helix influenced the membrane interactions of the CYP 17A1 globular domain. In some simulations, the mutated TM-helix obstructed the substrate access tunnel from the membrane to the CYP active site, indicating a possible effect on enzyme function.

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“…A number of studies have been established on the characteristic features of P450s in higher mammals including man and a model insect Drosophila melanogaster. These features expedite the identification of these cytochrome P450 enzymes and which among others include; the access paths for substrates and products and the distinctive properties of the active sites with web of water molecules [15]. However, detailed studies on the features of Cytochrome P450s in A. gambiae and more significantly their regional distribution which are quintessential for strategic planning of vector control programmes across three different regions of the African continent are scanty.…”

Section: Introductionmentioning

confidence: 99%

Cytochrome P450s in Anopheles gambiae (Diptera: Culicidae) and Insecticide Resistance in Africa: A Mini Review

Mohammed¹,

Mailafia²,

Malang³

et al. 2017

Entomol Ornithol Herpetol

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Cytochrome P450s are known to be critical for the detoxification and/or activation of xenobiotics such as insecticides in all living organisms including Anopheles gambiae. Many studies have demonstrated the role of P450s in insecticide resistance in A. gambiae. However, little is known about the impact of distribution in the African subcontinent. In this paper therefore, we analyse the P450 clans, the CYP6 family, localisation and function of A. gambiae CYPs, their insecticide substrates, regional distribution in the African continent and their role in insecticide resistance. This investigation from published data revealed that in the Central region; CYP6Z3, CYP6Z1, CYP12F2, CYP6P4, CYP6GA1, CYP6Z3 (Yaoundé, Cameroun) have bendiocarb, DDT and pyrethroids as substrates; in the Eastern region: CYP314A1 and CYP12F1 (Tanzania and Zanzibar) have DDT as a substrate, CYP32A3, CYP6Z1 and CYP6Z2 (Western Kenya) have DDT, carbaryl and permethrin; whilst in the Western region: CYP6AG1, CYP6Z2, CYP6Z3, CY6P3, CYP6P4, CYP6M2 (Ghana), CYP6M2, CYP6P3 (Benin), CYP325A3, CYP6P3 and CYP6M2 (Nigeria) all have DDT, carbaryl, permethrin, trans-and cis-permethrin, deltamethrin, bendiocarb as substrates. Additionally, CYP6M2, CYP6P3, CYP6Z3 (Côte d'Ivoire), CYP6P3, CYP6Z2 and CYP9J5 (Burkina Faso) have bendiocarb, DDT plus pyrethroids and only pyrethroids as substrates respectively. Interestingly, CYP6P3 is observed to metabolize all the available insecticides (DDT, pyrethroid, trans-and cis-permethrin, deltamethrin and bendiocarb), indicating possible insecticide cross resistance across all the three regions of Africa. A more detailed understanding of the substrate specificities of various P450s and the geographical distribution of insecticide resistance in Africa is quintessential for an effective resistance management.

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Structural features and dynamic investigations of the membrane-bound cytochrome P450 17A1

Cited by 18 publications

References 53 publications

Lipid molecules can induce an opening of membrane-facing tunnels in cytochrome P450 1A2

Lipid molecules can induce an opening of membrane-facing tunnels in cytochrome P450 1A2

Influence of Transmembrane Helix Mutations on Cytochrome P450-Membrane Interactions and Function

Cytochrome P450s in Anopheles gambiae (Diptera: Culicidae) and Insecticide Resistance in Africa: A Mini Review

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