Further Characterization of Hemopressin Peptide Fragments in the Opioid and Cannabinoid Systems

Szlávicz, Eszter; Perera, Pannilage Shiromi; Tömböly, Csaba; Helyes, Zsuzsanna; Zádor, Ferenc; Benyhe, Sándor; Borsodi, Anna; Bojnik, Engin

doi:10.1213/ane.0000000000000964

Cited by 11 publications

(3 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In our functional [ 35 S]GTPγS binding experiments, Hp(1-7) and Hp(1-9) behaved as very weak agonists (if at all), and could not stimulate [ 35 S]GTPγS binding significantly. Recently, a very similar [ 35 S]GTPγS stimulatory effect was observed in competitive radioligand and [ 35 S]GTPγS binding studies (Szlavicz et al, 2015). It was found that Hp(1-7) and Hp(1-9) slightly activated G-proteins in a naloxone-sensitive manner and that the peptides directly interacted with the CB1 and MOP receptors as well.…”

Section: Discussionsupporting

confidence: 53%

Investigation of receptor binding and functional characteristics of hemopressin(1–7)

et al. 2016

View full text Add to dashboard Cite

The orally active, α-hemoglobin derived hemopressin (PVNFKFLSH, Hp(1-9)) and its truncated (PVNFKFL, Hp(1-7) and PVNFKF, Hp(1-6)) and extended ((R)VDPVNFKFLSH, VD-Hp(1-9) and RVD-Hp(1-9)) derivatives have been postulated to be the endogenous peptide ligands of the cannabinoid receptor type 1 (CB1). In an attempt to create a versatile peptidic research tool for the direct study of the CB1 receptor-peptide ligand interactions, Hp(1-7) was radiolabeled and in vitro characterized in rat and CB1 knockout mouse brain membrane homogenates. In saturation and competition radioligand binding studies, [(3)H]Hp(1-7) labeled membrane receptors with high densities and displayed specific binding to a receptor protein, but seemingly not to the cannabinoid type 1, in comparison the results with the prototypic JWH-018, AM251, rimonabant, Hp(1-9) and RVD-Hp(1-9) (pepcan 12) ligands in both rat brain and CB1 knockout mouse brain homogenates. Furthermore, functional [(35)S]GTP γS binding studies revealed that Hp(1-7) and Hp(1-9) only weakly activated G-proteins in both brain membrane homogenates. Based on our findings and the latest literature data, we assume that the Hp(1-7) peptide fragment may be an allosteric ligand or indirect regulator of the endocannabinoid system rather than an endogenous ligand of the CB1 receptor.

show abstract

Section: Discussionsupporting

confidence: 53%

Investigation of receptor binding and functional characteristics of hemopressin(1–7)

et al. 2016

View full text Add to dashboard Cite

show abstract

“…The human variant of Hp (PVNFKLLSH) has a mutation in position 6, where a Leucine substitutes the second Phenylalanine. 4 Hemopressin proved to exert an antinociceptive action; [5][6][7] it was reported to cause dose-dependent hypotension in rats, rabbits, and mice [8][9][10] and to induce anxiogenic-like effects in rats. 11 In addition, hemopressin was shown to inhibit food intake in normal and obese animal models without causing obvious adverse side effects.…”

Section: Introductionmentioning

confidence: 99%

DOES hemopressin bind metal ions in vivo?

et al. 2016

View full text Add to dashboard Cite

Hemopressin is a neuropeptide, derived from the degradation of the α(1)-chain of hemoglobin, and possesses several pharmacologic properties, such as the ability to block cannabinoid CB1 receptor activity, to cause dose-dependent hypotension and to inhibit food intake. Actually, human hemopressin (PVNFKLLSH) is only the precursor of a class of longer peptides, called "Pepcans", which bear additional residues at their amino-terminus and possess slightly different chemical and biological properties with respect to hemopressin. The presence of a histidyl residue and the free terminal amine imparts to hemopressin and its derivatives good binding properties towards transition metal ions. In this paper, we present a wide investigation on the complex-formation equilibria of human hemopressin and three analogues towards the Cu(ii) and Ni(ii) ions. The study showed that the main coordination site is always the amino terminus (if not protected), while the C-terminal histidine acts only as an anchoring site for the metal ions at acidic pH, with the formation of a macrochelate complex. The presence of additional residues in N-terminal position produces significant differences in the protonation and complex-formation behaviors of these peptides, which can be explained in terms of charge of the ligand and coordination environment. Although the participation of metal ions in the biological activity of hemopressin and Pepcans has not yet been demonstrated, the data reported here can help to shed light on the mechanisms governing the action of these neuropeptides in vivo.

show abstract

“…Neste mesmo estudo, constatou-se a seletividade da hemopressina pelos receptores canabinoides CB1, por meio da utilização de anticorpos sensíveis ao estado de conformação para diversos receptores acoplados a proteína G, dentre eles, os receptores opioides e canabinoides, demonstrando atividade antagonista ou agonista inversa aos receptores CB1, uma vez que foi capaz de bloquear a atividade constitutiva destes receptores como o antagonista rimonabanto. Além da ativação dos receptores CB1, a hemopressina (PVNFKFLSH) e o seu fragmento C-terminal truncado hemopressina 1-7 (PVNFKFL), mostraram leve efeito modulador no sistema opioide (SZLAVICZ et al, 2015). Também no estudo de ligação e funcionalização do fragmento peptídico hemopressina marcada radioativamente (1-7) (PVNFKFL) mostrou ligação independente de CB1 em cérebro de rato tipo selvagem e a má ativação das proteínas G.…”

Section: Hemopressina E Suas Propriedades Farmacológicasunclassified

Produção e purificação do peptídeo hemopressina visando potencial farmacológico

Oliveira¹

View full text Add to dashboard Cite

Hemopressin is a nonapeptide derived from the α-1 chain of mammalian hemoglobin known for its hypotensive, analgesic and appetite suppressive effects. This peptide acts as a CB1 cannabinoid receptor inverse agonist or antagonist. Dysfunctions in the endocannabinoid system are associated with some pathologies and morbidity, such as obesity, neuropathic pain, depression, inflammation, Alzheimer and Parkinson diseases.This study was focused on the production and purification of synthetic and recombinant hemopressin, aiming future evaluation of their biological properties. To obtain the recombinant peptide, the nucleotide sequence encoding hemopressin was inserted into the expression vector pGEX-4T-1 (GE Healthcare) fused to the tag-GST and expressed in Escherichia coli BL21-CodonPlus. Several expression conditions were tested to determine the best expression parameters. Solid-phase synthesis of human and murine hemopressin was performed using the Fmoc/t-butyl strategy. The amino acid sequences of both forms of hemopressin was confirmed by MALDI mass spectrometry in the LIFT TM mode at an Autoflex speed equipment (BrukerDaltonics, Bilerica, USA).Purification of the synthetic peptide was performed by reversed-phase chromatography (Shimadzu LC-20AT) on a C18 semi-preparative column (Jupiter 5 µm 300 Å). Mass spectrometric analysis of human and murine hemopressins showed synthetic peptides with a molecular mass of 1,053.6 Da and 1,087.6 Da, respectively. The molecular mass of murine recombinant hemopressin fused to GST was approximately 31 kDa, a value that was obtained by SDS-PAGE analysis and confirmed by Western Blotting. The expression yield was evaluated by densitometry, which showed an equivalent expression yield of 147,8 mg/L of the recombinant protein.

show abstract

Further Characterization of Hemopressin Peptide Fragments in the Opioid and Cannabinoid Systems

Abstract: Here, we further confirm that hemopressins can modulate CB1 receptors and can have a slight modulatory effect on the opioid system.

Cited by 11 publications

References 28 publications

Investigation of receptor binding and functional characteristics of hemopressin(1–7)

Investigation of receptor binding and functional characteristics of hemopressin(1–7)

DOES hemopressin bind metal ions in vivo?

Produção e purificação do peptídeo hemopressina visando potencial farmacológico

Contact Info

Product

Resources

About