Establishment of a steroid binding assay for membrane progesterone receptor alpha (PAQR7) by using graphene quantum dots (GQDs)

Jyoti, Md. Maisum Sarwar; Rana, Md. Rubel; Ali, Md. Hasan; Tokumoto, Toshinobu

doi:10.1016/j.bbrc.2022.01.002

Cited by 9 publications

(11 citation statements)

References 38 publications

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“…A critical criterion for receptor designation is the presence of specific, high-affinity ligand binding. This criterion has been met by different laboratories for mPRs in prokaryotic ( Escherichia coli ), yeast, numerous mammalian expression systems, and most recently with purified recombinant mPRα coupled to graphene quantum dots [ 1 , 3 , 8 , 20 , 28 ]. Long-term culture of mammalian cells transfected with mPRs is required to obtain sufficient expression of the protein on the plasma membrane to detect progesterone binding [ 2 ].…”

Section: Ligand Bindingmentioning

confidence: 99%

Membrane Progesterone Receptors (mPRs, PAQRs): Review of Structural and Signaling Characteristics

Thomas

2022

Cells

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The role of membrane progesterone receptors (mPRs), which belong to the progestin and adipoQ receptor (PAQR) family, in mediating rapid, nongenomic (non-classical) progestogen actions has been extensively studied since their identification 20 years ago. Although the mPRs have been implicated in progestogen regulation of numerous reproductive and non-reproductive functions in vertebrates, several critical aspects of their structure and signaling functions have been unresolved until recently and remain the subject of considerable debate. This paper briefly reviews recent developments in our understanding of the structure and functional characteristics of mPRs. The proposed membrane topology of mPRα, the structure of its ligand-binding site, and the binding affinities of steroids were predicted from homology modeling based on the structures of other PAQRs, adiponectin receptors, and confirmed by mutational analysis and ligand-binding assays. Extensive data demonstrating that mPR-dependent progestogen regulation of intracellular signaling through mPRs is mediated by activation of G proteins are reviewed. Close association of mPRα with progesterone membrane receptor component 1 (PGRMC1), its role as an adaptor protein to mediate cell-surface expression of mPRα and mPRα-dependent progestogen signaling has been demonstrated in several vertebrate models. In addition, evidence is presented that mPRs can regulate the activity of other hormone receptors.

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Section: Ligand Bindingmentioning

confidence: 99%

Membrane Progesterone Receptors (mPRs, PAQRs): Review of Structural and Signaling Characteristics

Thomas

2022

Cells

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“…The binding properties of 2-HPA and its analogues to mPRα were evaluated by GQD-hmPRα according to the method described previously 21 .…”

Section: Methodsmentioning

confidence: 99%

“…We have also succeeded in expressing and purifying mPRα molecules using yeast, and have developed a high-throughput screening method for mPR-responsive substances using GQD-mPRα, which is a nanoparticle-associated mPRα 20,21 . On the other hand, we are also trying to isolate and purify mPR-reactive natural products using these screening methods 22 .…”

Section: Introductionmentioning

confidence: 99%

“…We have developed in vivo zebrafish oocyte maturation and ovulation induction methods as screening methods for mPR-responsive substances, as well as a test method using the modified luciferase gene (Glosensor) to measure intracellular cAMP concentration 18,19 . We have also succeeded in expressing and purifying mPRα molecules using yeast, and have developed a high-throughput screening method for mPR-responsive substances using GQD-mPRα, which is a nanoparticle-associated mPRα 20,21 . On the other hand, we are also trying to isolate and purify mPR-reactive natural products using these screening methods 22 .…”

Section: Introductionmentioning

confidence: 99%

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Discovery of specific activity of 2-HPA acting on the membrane progestin receptor α (paqr7) by purification of natural products from the marine algaePadina

Amin,

Acharjee,

Sarwar Jyoti

et al. 2023

Preprint

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Membrane progestin receptors (mPRs) are members of the progestin and adipoQ (PAQR) receptor family that are stimulated by endogenous steroids to initiate rapid intracellular signalling through a nongenomic pathway. Previously, water-soluble compounds with mPRα-binding activity from the marine algaePadina arborescenswere fractionated by HPLC steps. In this study, the structure of one of the major compounds in the fraction was identified as 2-hydroxypentanoic acid (2-HPA) using Nuclear Magnetic Resonance spectroscopy. 2-HPA showed a substantial competitive binding affinity for hmPRα in the GQD-hmPRα binding assay. In contrast, synthetic structural analogues of 2-HPA showed no competitive binding activity. The physiological activity of 2-HPA and its analogues was then investigated usingin vitrogoldfish andin vivozebrafish oocyte maturation and ovulation assays. As with the hmPRα binding assay, only 2-HPA showed inhibitory activity on oocyte maturation and ovulation of fish oocytes. Furthermore, the inhibitory activity of 2-HPA was compared between S- and R-type 2-HPA. The results showed that both types had the same level of activity. These results indicate that 2-HPA, found as a secreted compound fromPadina arborescens, is a novel mPRα antagonist and its chemical structure is highly restricted to show its activity.

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“…We have developed in vivo zebra sh oocyte maturation and ovulation induction methods as screening methods for mPR-responsive substances, as well as a test method using the modi ed luciferase gene (Glosensor) to measure intracellular cAMP concentration 18,19 . We have also succeeded in expressing and purifying mPRa molecules using yeast, and have developed a high-throughput screening method for mPR-responsive substances using GQD-mPRa, which is a nanoparticle-associated mPRa 20,21 . On the other hand, we are also trying to isolate and purify mPR-reactive natural products using these screening methods 22 .…”

mentioning

confidence: 99%

Discovery of specific activity of 2-HPA acting on the membrane progestin receptor alpha (paqr7) by purification of natural products from the marine algae Padina

Amin,

Acharjee,

Jyoti

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Membrane progestin receptors (mPRs) are members of the progestin and adipoQ (PAQR) receptor family that are stimulated by endogenous steroids to initiate rapid intracellular signalling through a nongenomic pathway. Previously, water-soluble compounds with mPRα-binding activity from the marine algae Padina arborescens were fractionated by HPLC steps. In this study, the structure of one of the major compounds in the fraction was identified as 2-hydroxypentanoic acid (2-HPA) using Nuclear Magnetic Resonance spectroscopy. 2-HPA showed a substantial competitive binding affinity for hmPRα in the GQD-hmPRα binding assay. In contrast, synthetic structural analogues of 2-HPA showed no competitive binding activity. The physiological activity of 2-HPA and its analogues was then investigated using in vitro goldfish and in vivo zebrafish oocyte maturation and ovulation assays. As with the hmPRα binding assay, only 2-HPA showed inhibitory activity on oocyte maturation and ovulation of fish oocytes. Furthermore, the inhibitory activity of 2-HPA was compared between S- and R-type 2-HPA. The results showed that both types had the same level of activity. These results indicate that 2-HPA, found as a secreted compound from Padina arborescens, is a novel mPRα antagonist and its chemical structure is highly restricted to show its activity.

show abstract

Establishment of a steroid binding assay for membrane progesterone receptor alpha (PAQR7) by using graphene quantum dots (GQDs)

Cited by 9 publications

References 38 publications

Membrane Progesterone Receptors (mPRs, PAQRs): Review of Structural and Signaling Characteristics

Membrane Progesterone Receptors (mPRs, PAQRs): Review of Structural and Signaling Characteristics

Discovery of specific activity of 2-HPA acting on the membrane progestin receptor α (paqr7) by purification of natural products from the marine algaePadina

Discovery of specific activity of 2-HPA acting on the membrane progestin receptor alpha (paqr7) by purification of natural products from the marine algae Padina

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