Purification and Cloning of Opioid Receptors

Ly, Li; Su, Yu-Feng; Zm, Zhang; Cs, Wong; Kj, Chang

doi:10.1037/e495942006-012

Cited by 3 publications

(3 citation statements)

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“…Three types of opioid receptors have been characterized and cloned in rats, mice and humans and designated as δ, µ, and κ [1][2][3]. The endogenous opioids are classified into β-endorphins, met-enkephalins and dynorphins based on their precursors: pro-opiomelanocortin, pro-enkephalin and pro-dynorphin [4][5][6].…”

Section: Endogenous Opioid Systemmentioning

confidence: 99%

The Involvement of the Nitric Oxide in the Effects and Expression of Opioid Receptors During Peripheral Inflammation

Pol

2007

CMC

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Peripheral inflammation enhances the antinociceptive effects of opioid receptor agonists through the activation of peripheral opioid receptors whose expression also increases during inflammatory pain. Similarly, intestinal inflammation also increases the antitransit and antiexudative effects of opioids as well as the expression of neuronal and extra-neuronal opioid receptors located in the gut. Nitric oxide has been described either as pro- or antiinflammatory and could produce both pro- and antinociceptive effects. In addition, numerous studies have shown that the L-arginine-nitric oxide-cGMP system participates in the antinociceptive and in the intestinal effects produced by opioids during peripheral inflammation by enhancing their effects. Thus, substances capable of inhibiting cyclic guanosine-3',5'-monophosphate (cGMP) degradation or nitric oxide donors increase the analgesic effects of opioid receptor agonists during peripheral inflammation. At the same time, the administration of nitric oxide synthase (NOS) or guanylate cyclase inhibitors decreases those effects. In accordance with these results, different clinical trials have also demonstrated that the co-administration of nitric oxide donors with opioids is highly beneficial in the treatment of pain in patients. In the gut, nitric oxide has a further pro- and antiinflammatory action. It is also involved in the enhanced antitransit and antiexudative effects produced by opioids and in the up-regulation of the mu-opioid receptor gene transcription observed in the inflamed intestine. To sum up, a better knowledge of the involvement of the L-arginine-nitric oxide-cGMP pathway in the opioid mechanisms of action and a better understanding of the pathways that regulate the expression of opioid receptors during peripheral inflammation are essential to developing improved analgesic/antiinflammatory therapies.

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Section: Endogenous Opioid Systemmentioning

confidence: 99%

The Involvement of the Nitric Oxide in the Effects and Expression of Opioid Receptors During Peripheral Inflammation

Pol

2007

CMC

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“…We also present a setup for the functional immobilization of GPCRs and demonstrate it for opioid receptors. Opioid receptors are known to be unstable in detergents [reviewed elsewhere (Simon, 1986(Simon, , 1991Smith and Loh, 1991;Li et al, 1993)] and we therefore sought an optimal balance between functionality (such as ligand binding) and harsher extraction conditions leading to homogeneity of the immobilized receptor. In order to characterize qualitatively the immobilized receptor, we compared the ligand binding profiles of immobilized and membrane-embedded receptor.…”

Section: Introductionmentioning

confidence: 99%

Engineering and functional immobilization of opioid receptors

Ott¹,

Neldner²,

Cèbe³

et al. 2005

Protein Engineering, Design and Selection

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Opioid receptors, like many G protein-coupled receptors (GPCRs), are notoriously unstable in detergents. We have now developed a more stable variant of the mu-opioid receptor (MOR) and also a method for the immobilization of solubilized, functional opioid receptors on a solid phase (magnetic beads). Starting with the intrinsically more stable kappa-opioid receptor (KOR), we optimized the conditions (i.e. detergents and stabilizing ligands) for receptor extraction from lipid bilayers of HEK293T cells to obtain maximal amounts of functional, immobilized receptor. After immobilization, the ligand binding profile remains the same as observed for the membrane-embedded receptor. For the immobilized wild-type mu-opioid receptor, however, no conditions were found under which ligand binding capacity was retained. To solve this problem, we engineered the receptor chimera KKM where the N-terminus and the first transmembrane helix (TM1) of wild-type MOR is exchanged for the homologous receptor parts of the wild-type KOR. This hybrid receptor behaves exactly as the wild-type MOR in functional assays. Interestingly, the modified MOR is expressed at six times higher levels than wild-type MOR and is similarly stable as wild-type KOR after immobilization. Hence the immobilized MOR, represented by the chimera KKM, is now also amenable for biophysical characterization. These results are encouraging for future stability engineering of GPCRs.

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“…Ƞ ȝ (ȝ-OR), Ƞ ț (ț-OR) (Li et al, 1993) țĮȚ Ƞ ORL-1 (opioid receptor-like 1) (Mollereau et al, 1994). ȅȚ ȠʌȚȠİȚįİȓȢ ȣʌȠįȠȤİȓȢ țȜȦȞȠʌȠȚȒșȘțĮȞ ıĲȚȢ ĮȡȤȑȢ ĲȘȢ įİțĮİĲȓĮȢ ĲȠȣ 1990 țĮȚ İțĲİȞİȓȢ ĳĮȡȝĮțȠȜȠȖȚțȑȢ ȝİȜȑĲİȢ ȤĮȡĮțĲȒȡȚıĮȞ (Snyder and Pasternak, 2003;Pasternak, 2005 (Kieffer and Gaveriaux-Ruff, 2002).…”

Section: Gȕȗ ȣʌƞȝƞȟȑįİȣmentioning

confidence: 99%

Ανάλυση Νέων Σηματοδοτικών Μονοπατιών Των Οπιοειδών Υποδοχέων Που Οδηγούν Σε Αλλαγές Της Συναπτοσωμικής Πλαστικότητας

Georganta¹,

Γεωργαντά²

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Οι οπιοειδείς υποδοχείς εμπλέκονται στην ανάπτυξη και διαφοροποίηση των νευρικών κυττάρων. Προηγούμενες μελέτες του εργαστηρίου μας έδειξαν ότι το συντηρημένο YXXL μοτίβο στο καρβοξυτελικό άκρο του μ-οπιοειδούς υποδοχέα (μ-OR) αποτελεί σημείο αγκυροβόλησης για τον μεταγραφικό παράγοντα STAT5A, ο οποίος φωσφορυλιώνεται μετά την ενεργοποίηση του μ-OR. Δεδομένου ότι και ο δ-οπιοειδής υποδοχέας (δ-OR) περιέχει το ίδιο δομικό μοτίβο στο καρβοξυτελικό του άκρο (δ-CT), αποδείξαμε ότι η STAT5Β αγκυροβολεί σε αυτή την περιοχή του δ-CT. Έκθεση των HEK293 κυττάρων, που εκφράζουν τον flag-δ-OR, σε δ-οπιοειδείς αγωνιστές οδηγεί στη Gαi/o-εξαρτώμενη φωσφορυλίωση της STAT5Β μέσω μιας c-Src τυροσινικής κινάσης. Επιπλέον μελέτες έδειξαν ότι ο δ-OR λειτουργεί ως πλατφόρμα για το σχηματισμό ενός πολύ-πρωτεϊνικού συμπλόκου, αποτελούμενου από τη STAT5B, τη c-Src κινάση, τις Gβγ και επιλεκτικές Gαi/o υπομονάδες των G πρωτεϊνών. Επιπρόσθετα, τα αποτελέσματά μας έδειξαν ότι η ενεργοποίηση του δ-OR με το οπιοειδές πεπτίδιο DSLET συμβάλλει στην επιβίωση νευρικών κυττάρων (Neuro-2A και SH-SY5Y) και επιπλέον επάγει τη νευριτική ανάπτυξη αυτών, μέσω ενός μονοπατιού που περιλαμβάνει τις Gαi/o υπομονάδες και τη φωσφορυλιωμένη STAT5Β. Τα δεδομένα αυτά αποδεικνύουν ότι το σηματοδοτικό μονοπάτι του δ-OR που οδηγεί στην ενεργοποίηση της STAT5Β αποτελεί ένα μηχανισμό μέσω του οποίου επιτυγχάνεται η επιβίωση και διαφοροποίηση των νευρικών κυττάρων.

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Purification and Cloning of Opioid Receptors

Cited by 3 publications

References 0 publications

The Involvement of the Nitric Oxide in the Effects and Expression of Opioid Receptors During Peripheral Inflammation

The Involvement of the Nitric Oxide in the Effects and Expression of Opioid Receptors During Peripheral Inflammation

Engineering and functional immobilization of opioid receptors

Ανάλυση Νέων Σηματοδοτικών Μονοπατιών Των Οπιοειδών Υποδοχέων Που Οδηγούν Σε Αλλαγές Της Συναπτοσωμικής Πλαστικότητας

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