Advances in the Understanding of Protein-Protein Interactions in Drug Metabolizing Enzymes through the Use of Biophysical Techniques

Lampe, Jed N.

doi:10.3389/fphar.2017.00521

Cited by 15 publications

(8 citation statements)

References 123 publications

(170 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Both Cyp1a2 and Cyp2e1 proteins bind Pgrmc1, and the proteins are expressed at lower levels in Pgrmc1 KO liver. PGRMC1 most likely controls enzyme activity by stabilizing cytochromes P450 rather than affecting the catalytic cycle of the enzyme, but a concomitant effect on the k cat of cytochromes P450 upon PGRMC1 binding cannot be ruled out as protein–protein interactions are also known to alter cytochrome P450 activity ( 39 , 40 , 41 , 42 ). Evidence exists that PGRMC1 binds cytochromes P450 directly.…”

Section: Discussionmentioning

confidence: 99%

Progesterone receptor membrane component 1 (PGRMC1) binds and stabilizes cytochromes P450 through a heme-independent mechanism

McGuire

Mukhopadhyay

Myers

et al. 2021

Journal of Biological Chemistry

View full text Add to dashboard Cite

This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

show abstract

Section: Discussionmentioning

confidence: 99%

Progesterone receptor membrane component 1 (PGRMC1) binds and stabilizes cytochromes P450 through a heme-independent mechanism

McGuire

Mukhopadhyay

Myers

et al. 2021

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Allosteric receptor-receptor interaction, which is still an underexplored mechanism of receptor modulation [37], has been suggested to be an especially effective approach to control NMDA receptor function [38][39][40]. The functional readout from the interaction between two proteins is often modified by small molecules [41,42], and it is well established that ouabain binding to the potassium-binding state of the catalytic NKA subunit changes its conformation [43][44][45]. Lack of effect of the low ouabain concentration on membrane potential (Supp.…”

Section: Discussionmentioning

confidence: 99%

Ouabain Modulates the Functional Interaction Between Na,K-ATPase and NMDA Receptor

et al. 2020

View full text Add to dashboard Cite

The N-methyl-D-aspartate (NMDA) receptor plays an essential role in glutamatergic transmission and synaptic plasticity and researchers are seeking for different modulators of NMDA receptor function. One possible mechanism for its regulation could be through adjacent membrane proteins. NMDA receptors coprecipitate with Na,K-ATPase, indicating a potential interaction of these two proteins. Ouabain, a mammalian cardiotonic steroid that specifically binds to Na,K-ATPase and affects its conformation, can protect from some toxic effects of NMDA receptor activation. Here we have examined whether NMDA receptor activity and downstream effects can be modulated by physiological ouabain concentrations. The spatial colocalization between NMDA receptors and the Na,K-ATPase catalytic subunits on dendrites of cultured rat hippocampal neurons was analyzed with superresolution dSTORM microscopy. The functional interaction was analyzed with calcium imaging of single hippocampal neurons exposed to 10 μM NMDA in presence and absence of ouabain and by determination of the ouabain effect on NMDA receptordependent long-term potentiation. We show that NMDA receptors and the Na,K-ATPase catalytic subunits alpha1 and alpha3 exist in same protein complex and that ouabain in nanomolar concentration consistently reduces the calcium response to NMDA. Downregulation of the NMDA response is not associated with internalization of the receptor or with alterations in its state of Src phosphorylation. Ouabain in nanomolar concentration elicits a long-term potentiation response. Our findings suggest that ouabain binding to a fraction of Na,K-ATPase molecules that cluster with the NMDA receptors will, via a conformational effect on the NMDA receptors, cause moderate but consistent reduction of NMDA receptor response at synaptic activation. Evgeny E. Akkuratov and Linda Westin contributed equally to this work.

show abstract

“…Identifying protein interaction sites and uncovering the interaction mechanism is closely relevant in the drug development industry. In addition, unveiling of protein interaction partners allows biologists to construct protein interaction networks, which in turn facilitate the understanding of many biological and clinical observations [ 19 , 20 , 21 , 22 ]. The data in this study indicated that Tb -DdlA could not maintain its activity long in vitro under the conditions tested, which might be attributable to its synergistic effect with potential interacting proteins that stabilized its activity in vivo.…”

Section: Discussionmentioning

confidence: 99%

The Biological Properties and Potential Interacting Proteins of d-Alanyl-d-alanine Ligase A from Mycobacterium tuberculosis

Yang

Wang

et al. 2018

Molecules

View full text Add to dashboard Cite

(1) Background: d-alanine-d-alanine ligase (DdlA), an effective target for drug development to combat against Mycobacterium tuberculosis (Mtb), which threatens human health globally, supplies a substrate of d-alanyl-d-alanine for peptidoglycan crosslinking by catalyzing the dimerization of two d-alanines. To obtain a better understanding of DdlA profiles and develop a colorimetric assay for high-throughput inhibitor screening, we focused on explicating and characterizing Tb-DdlA. (2) Methods and Results: Rv2981c (ddlA) was expressed in Escherichia coli, and the purified Tb-DdlA was identified using (anti)-polyhistidine antibody followed by DdlA activity confirmation by measuring the released orthophosphate via colorimetric assay and the yielded d-alanyl-d-alanine through high performance thin layer chromatography (HP-TLC). The kinetic assays on Tb-DdlA indicated that Tb-DdlA exhibited a higher affinity to ATP (KmATP: 50.327 ± 4.652 μmol/L) than alanine (KmAla: 1.011 ± 0.094 mmol/L). A colorimetric assay for Tb-DdlA activity was developed for high-throughput screening of DdlA inhibitors in this study. In addition, we presented an analysis on Tb-DdlA interaction partners by pull-down assay and MS/MS. Eight putative interaction partners of Tb-DdlA were identified. (3) Conclusions: Our dataset provided a valuable resource for exploring Tb-DdlA biology, and developed an easy colorimetric assay for screening of Tb-DdlA inhibitors.

show abstract

Advances in the Understanding of Protein-Protein Interactions in Drug Metabolizing Enzymes through the Use of Biophysical Techniques

Cited by 15 publications

References 123 publications

Progesterone receptor membrane component 1 (PGRMC1) binds and stabilizes cytochromes P450 through a heme-independent mechanism

Progesterone receptor membrane component 1 (PGRMC1) binds and stabilizes cytochromes P450 through a heme-independent mechanism

Ouabain Modulates the Functional Interaction Between Na,K-ATPase and NMDA Receptor

The Biological Properties and Potential Interacting Proteins of d-Alanyl-d-alanine Ligase A from Mycobacterium tuberculosis

Contact Info

Product

Resources

About