Methionine Proximity Assay, a Novel Method for Exploring Peptide Ligand–receptor Interaction

Řihakova, Lenka; Deraët, Maud; Auger‐Messier, Mannix; Pérodin, Jacqueline; Boucard, Antony A.; Guillemette, Gaétan; Leduc, Richard; Lavigne, Pierre; Escher, Emanuel

doi:10.1081/rrs-120014603

Cited by 42 publications

(47 citation statements)

References 27 publications

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“…A bias for recovery of peptide fragments from these TM segments was not observed for unmodified peptide fragments. The propensity of benzophenone-type photoligands to react with methionine residues (Rihakova et al, 2002) is reflected by the fact that highest labeling is observed in methionine-containing sequences Gln169-Met170 in TM3, Leu268-Met269-Ile270 in TM5, and Met295-Met296-Tyr297 in TM6.…”

Section: Resultsmentioning

confidence: 99%

A Three-Dimensional Model for the Substrate Binding Domain of the Multidrug ATP Binding Cassette Transporter LmrA

Ecker

Pleban²,

Kopp³

et al. 2004

Mol Pharmacol

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Multidrug resistance presents a major obstacle to the treatment of infectious diseases and cancer. LmrA, a bacterial ATP-dependent multidrug transporter, mediates efflux of hydrophobic cationic substrates, including antibiotics. The substrate-binding domain of LmrA was identified by using photo-affinity ligands, proteolytic degradation of LmrA, and identification of ligand-modified peptide fragments with matrix-assisted laser desorption ionization/time of flight mass spectrometry. In the nonenergized state, labeling occurred in the ␣-helical transmembrane segments (TM) 3, 5 and 6 of the membrane-spanning domain. Upon nucleotide binding, the accessibility of TM5 for substrates increased, whereas that of TM6 decreased. Inverse changes were observed upon ATP-hydrolysis. An atomic-detail model of dimeric LmrA was generated based on the template structure of the homologous transporter MsbA from Vibrio cholerae, allowing a three-dimensional visualization of the substrate-binding domain. Labeling of TM3 of one monomer occurred in a predicted area of contact with TM5 or TM6 of the opposite monomer, indicating substrate-binding at the monomer/monomer interface. Inverse changes in the reactivity of TM segments 5 and 6 suggest that substrate binding and release involves a repositioning of these helices during the catalytic cycle.

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Section: Resultsmentioning

confidence: 99%

A Three-Dimensional Model for the Substrate Binding Domain of the Multidrug ATP Binding Cassette Transporter LmrA

Ecker

Pleban²,

Kopp³

et al. 2004

Mol Pharmacol

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“…Benzophenonetype ligands have initially been reported to be nonselective (Dorman and Prestwich, 2000). A preferred reaction with methionine residues in proteins has been reported (Rihakova et al, 2002;Wahlstrom et al, 2002). Methionine residues in Fig.…”

Section: Resultsmentioning

confidence: 99%

P-Glycoprotein Substrate Binding Domains Are Located at the Transmembrane Domain/Transmembrane Domain Interfaces: A Combined Photoaffinity Labeling-Protein Homology Modeling Approach

Pleban¹,

Kopp²,

Csaszar³

et al. 2004

Mol Pharmacol

121

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P-glycoprotein (P-gp) is an energy-dependent multidrug efflux pump conferring resistance to cancer chemotherapy. Characterization of the mechanism of drug transport at a molecular level represents an important prerequisite for the design of pump inhibitors, which resensitize cancer cells to standard chemotherapy. In addition, P-glycoprotein plays an important role for early absorption, distribution, metabolism, excretion, and toxicity profiling in drug development. A set of propafenonetype substrate photoaffinity ligands has been used in this study in conjunction with matrix-assisted laser desorption/ionization timeof-flight mass spectrometry to define the substrate binding domain(s) of P-gp in more detail. The highest labeling was observed in transmembrane segments 3, 5, 8, and 11. A homology model for P-gp was generated on the basis of the dimeric crystal structure of Vibrio cholerae MsbA, an essential lipid transporter. Thereafter, the labeling pattern was projected onto the 3D atomic-detail model of P-gp to allow a visualization of the binding domain(s). Labeling is predicted by the model to occur at the two transmembrane domain/transmembrane domain interfaces formed between the amino-and carboxyl-terminal half of P-gp. These interfaces are formed by transmembrane (TM) segments 3 and 11 on one hand and TM segments 5 and 8 on the other hand. Available data on LmrA and AcrB, two bacterial multidrug efflux pumps, suggest that binding at domain interfaces may be a general feature of polyspecific drug efflux pumps.Multidrug resistance represents a serious obstacle to successful cancer chemotherapy. Although multifactorial in etiology, one type of multidrug resistance is associated with the overexpression of energy-dependent membrane-bound pumps, which intercept and efflux drugs before they reach their intracellular target structures. P-glycoprotein (ABCB1) represents a paradigm ATP-dependent efflux pump expressed in human cancer cells. In addition to its expression in cancer cells, P-gp is also physiologically expressed in a number of tissues such as intestinal epithelial cells, at the brush border of renal tubule epithelial cells, the canalicular side of hepatocytes, and in capillary endothelial cells forming the blood-brain barrier. It thus interferes with oral drug absorption and drug delivery to the brain, and it enhances renal and biliary excretion. P-gp has therefore attracted considerable attention as a nontarget in the field of drug development, because for a large number of active compounds, interaction with P-glycoprotein might compromise their future development into a drug. Considerable energy has therefore been devoted to the characterization of molecular features that make compounds P-gp substrates and to the definition of the molecular mechanism of drug transport by P-gp. A number of studies have dealt with the kinetics and thermodynamics of the transport process

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“…1c) (21,(23)(24)(25). In the FIGURE 1. a, MPA (59). Following the binding of the Bpa-substituted ligand with its target, the benzophenone group of the Bpa photoprobe is photoactivated at 350 -360 nm to generate a reactive triplet biradical ketone intermediate (19).…”

Section: Methodsmentioning

confidence: 99%

Structure of the Human Angiotensin II Type 1 (AT1) Receptor Bound to Angiotensin II from Multiple Chemoselective Photoprobe Contacts Reveals a Unique Peptide Binding Mode

Fillion

Cabana

Guillemette

et al. 2013

Journal of Biological Chemistry

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Background: G protein-coupled receptors (GPCRs) modulate a plethora of physiological processes. Results: The determination of 38 ligand/receptor contacts enabled the evidence-based modeling of a GPCR structure. Conclusion: The proposed GPCR structure assumes four interaction clusters with its ligand, which adopts a vertical binding mode. Significance: Elucidating the structure of GPCRs is central to their understanding and has several implications, e.g. rational drug design.

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Methionine Proximity Assay, a Novel Method for Exploring Peptide Ligand–receptor Interaction

Cited by 42 publications

References 27 publications

A Three-Dimensional Model for the Substrate Binding Domain of the Multidrug ATP Binding Cassette Transporter LmrA

A Three-Dimensional Model for the Substrate Binding Domain of the Multidrug ATP Binding Cassette Transporter LmrA

P-Glycoprotein Substrate Binding Domains Are Located at the Transmembrane Domain/Transmembrane Domain Interfaces: A Combined Photoaffinity Labeling-Protein Homology Modeling Approach

Structure of the Human Angiotensin II Type 1 (AT1) Receptor Bound to Angiotensin II from Multiple Chemoselective Photoprobe Contacts Reveals a Unique Peptide Binding Mode

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