Development of a novel ligand binding assay for relaxin family peptide receptor 3 and 4 using NanoLuc complementation.

Abstract:

:Relaxin family peptides perform a variety of biological functions by binding and activating relaxin family peptide receptor 1-4 (RXFP1-4), four A-class G protein-coupled receptors. In the present work, we developed a novel ligand binding assay for RXFP3 and RXFP4 based on NanoLuc complementation technology (NanoBiT). A synthetic ligation version of the low-affinity small complementation tag (SmBiT) was efficiently ligated to the A-chain N terminus of recombinant chimeric agonist R3/I5 using recombinant circular sortase A. After the ligation product R3/I5-SmBiT was mixed with human RXFP3 or RXFP4 genetically fused with a secretory large NanoLuc fragment (sLgBiT) at the N terminus, NanoLuc complementation was induced by high-affinity ligand-receptor binding. Binding kinetics and affinities of R3/I5-SmBiT with sLgBiT-fused RXFP3 and RXFP4 were conveniently measured according to the complementation-induced bioluminescence. Using R3/I5-SmBiT and the sLgBiT-fused receptor as a complementation pair, binding potencies of various ligands with RXFP3 and RXFP4 were quantitatively measured without the cumbersome washing step. The novel NanoBiT-based ligand binding assay is convenient for use and suitable for automation, thus will facilitate interaction studies of RXFP3 and RXFP4 with ligands in future. This assay can also be applied to some other plasma membrane receptors for pharmacological characterization of ligands in future studies.

journal_name

Amino Acids

journal_title

Amino acids

authors

Hu MJ,Shao XX,Li HZ,Nie WH,Wang JH,Liu YL,Xu ZG,Guo ZY

doi

10.1007/s00726-018-2588-5

subject

Has Abstract

pub_date

2018-08-01 00:00:00

pages

1111-1119

issue

8

eissn

0939-4451

issn

1438-2199

pii

10.1007/s00726-018-2588-5

journal_volume

50

pub_type

杂志文章
  • Synthesis and electrochemical behaviour of beta-halodehydroamino acid derivatives.

    abstract::Several new beta,beta-dihalo and beta-halo-beta-substituted dehydroalanines and dehydrodipeptides were synthesized by reacting the corresponding dehydroamino acid derivative with a N-halosuccinimide or in the case of beta,beta-di-iododehydroalanines with iodine. The results obtained confirmed that the stereochemical o...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-009-0466-x

    authors: Ferreira PM,Monteiro LS,Pereira G

    更新日期:2010-07-01 00:00:00

  • Expression of ABC-1 transporter is elevated in human glioma cells under irradiation and temozolomide treatment.

    abstract:OBJECTIVE:Chemo-therapeutic treatment of glioma patients has minor success. Little is known about mechanisms of a pronounced resistance of gliomas towards actual therapies, yet. ABC-1 belongs to the group of transporters known to be involved in the export of hydrophobic substances and vascular regulation. This study in...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-005-0171-3

    authors: Trog D,Moenkemann H,Haertel N,Schüller H,Golubnitschaja O

    更新日期:2005-03-01 00:00:00

  • The ubiquitin-proteasome system and autophagy are defective in the taurine-deficient heart.

    abstract::Taurine depletion leads to impaired mitochondrial function, as characterized by reduced ATP production and elevated superoxide generation. These defects can fundamentally alter cardiomyocyte function and if left unchanged can result in cell death. To protect against these stresses, cardiomyocytes possess quality contr...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-015-2053-7

    authors: Jong CJ,Ito T,Schaffer SW

    更新日期:2015-12-01 00:00:00

  • Hemolymph amino acid variations following behavioral and genetic changes in individual Drosophila larvae.

    abstract::This study investigated the effect of different sampling environments on hemolymph amino acid content of individual Drosophila melanogaster larvae. Hemolymph was collected from individual third instar larvae under cold-anesthetized, awake, and stress conditions. Qualitative and quantitative hemolymph amino acid analys...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-009-0284-1

    authors: Piyankarage SC,Augustin H,Featherstone DE,Shippy SA

    更新日期:2010-03-01 00:00:00

  • The gastro-intestinal tract as the major site of biological action of dietary melanoidins.

    abstract::Emerging evidence from laboratory researches has highlighted the bioactivity of food melanoidins and melanoproteins. Whilst such studies have been carried out with different in vitro systems, information about melanoidins absorption and bio-availability are scarce. However, they are generally considered as poorly abso...

    journal_title:Amino acids

    pub_type: 杂志文章,评审

    doi:10.1007/s00726-015-1951-z

    authors: Tagliazucchi D,Bellesia A

    更新日期:2015-06-01 00:00:00

  • Phenylalanine ammonia-lyase modified with polyethylene glycol: potential therapeutic agent for phenylketonuria.

    abstract::Phenylketonuria (PKU) is an autosomal recessive genetic disease caused by the defects in the phenylalanine hydroxylase (PAH) gene. Individuals homozygous for defective PAH alleles show elevated levels of systemic phenylalanine and should be under strict dietary control to reduce the risk of neuronal damage associated ...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-005-0218-5

    authors: Ikeda K,Schiltz E,Fujii T,Takahashi M,Mitsui K,Kodera Y,Matsushima A,Inada Y,Schulz GE,Nishimura H

    更新日期:2005-11-01 00:00:00

  • Taurine promotes connective tissue growth factor (CTGF) expression in osteoblasts through the ERK signal pathway.

    abstract::Taurine is found in bone tissue, but its function in skeletal tissue is not fully understood. The present study was undertaken to investigate regulation of gene expression of connective tissue growth factor (CTGF), and the roles of mitogen-activated protein kinases (MAPKs) in murine osteoblast MC3T3-E1 cells treated w...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-006-0380-4

    authors: Yuan LQ,Lu Y,Luo XH,Xie H,Wu XP,Liao EY

    更新日期:2007-01-01 00:00:00

  • The Schöllkopf chiron and transition metal mediated reactions, a powerful combination for stereoselective construction of cyclic alpha-quaternary-alpha-amino acid derivatives.

    abstract::The focus has been on the development of methodology for stereoselective preparation of spiroannulated intermediates of the Schöllkopf chiron and further transformations to cyclic alpha-amino acids. The spiroannulations are effected by Ru(II)-catalysed ring-closing metathesis reactions, by Ru(II)- and Pd(0)-catalysed ...

    journal_title:Amino acids

    pub_type: 杂志文章,评审

    doi:10.1007/s00726-007-0512-5

    authors: Undheim K

    更新日期:2008-04-01 00:00:00

  • Site-specific pegylation of an antimicrobial peptide increases resistance to Pseudomonas aeruginosa elastase.

    abstract::M33 is a branched peptide currently under preclinical characterization for the development of a new antibacterial drug against gram-negative bacteria. Here, we report its pegylation at the C-terminus of the three-lysine-branching core and the resulting increase in stability to Pseudomonas aeruginosa elastase. This pro...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-014-1686-2

    authors: Falciani C,Lozzi L,Scali S,Brunetti J,Bracci L,Pini A

    更新日期:2014-05-01 00:00:00

  • Synthesis and biological activity of oxytocin analogues containing unnatural amino acids in position 9: structure activity study.

    abstract::We report the solid phase synthesis and some pharmacological properties of 24 oxytocin (OT) analogues. Basic modifications at position 9 (introduction of L- or D-beta-(2-thienyl)-alanine [L- or D-Thi], or L- or D-3-Pyridylalanine [L- or D-3-Pal]) were combined with D-tyrosine(OEthyl) [D-Tyr(Et)] or D-1-naphthylalanine...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-009-0372-2

    authors: Magafa V,Borovicková L,Slaninová J,Cordopatis P

    更新日期:2010-05-01 00:00:00

  • A new method to synthesize creatine derivatives.

    abstract::Creatine is an amino acid that has a pivotal role in energy metabolism of cells. Creatine acts as an "ATP shuttle", carrying ATP to the sites where it is utilized, through its reversible phosphorylation by creatine kinase. Moreover, the creatine-phosphocreatine system delays ATP depletion during anoxia or ischemia, th...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-013-1525-x

    authors: Garbati P,Salis A,Adriano E,Galatini A,Damonte G,Balestrino M,Millo E

    更新日期:2013-10-01 00:00:00

  • Transglutaminases: future perspectives.

    abstract::This is the third special issue focused on "Transglutaminases" that is now available on this journal and dedicated to one of the pioneers of these enzymes, John Edward Folk, who died December 2010 [see in this issue Beninati et al. 2012a]. The first edition, "Polyamines and Transglutaminases" was published in Amino Ac...

    journal_title:Amino acids

    pub_type: 社论

    doi:10.1007/s00726-012-1431-7

    authors: Beninati S,Facchiano F,Piacentini M

    更新日期:2013-01-01 00:00:00

  • Design and synthesis of mono and bicyclic tetrapeptides thioester as potent inhibitor of histone deacetylases.

    abstract::Inhibitors of histone deacetylases (HDACs) are a promising class of anticancer agents that have an effect on gene regulation. The naturally occurring cyclic depsipeptide FK228 containing disulfide and Largazole possessing thioester functionalities act as pro-drugs and share the same HDAC inhibition mechanism in cell. ...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-014-1800-5

    authors: Hoque MA,Islam MS,Islam MN,Kato T,Nishino N,Ito A,Yoshida M

    更新日期:2014-10-01 00:00:00

  • Quantification of D-amino acids in human urine using GC-MS and HPLC.

    abstract::The relative amounts of free D-amino acids (D-AA) in the urine of seven healthy volunteers (age 27 to 49 years) were determined using chiral phase (Chirasil-L-Val) capillary gas chromatography in conjunction with selected ion monitoring mass spectrometry. The absolute amounts of free D-AA were determined by pre-column...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/BF00805848

    authors: Brückner H,Haasmann S,Friedrich A

    更新日期:1994-06-01 00:00:00

  • Isozymic nature of spore coat-associated alanine racemase of Bacillus subtilis.

    abstract::Spore coat-associated alanine racemase of Bacillus subtilis, which converts L-alanine to D-alanine, that is, the germinant to the competitive inhibitor, to regulate spore germination for survival of the organism under unfavorable growth conditions, was examined. The dormant spores, L-alanine-initiated germination of w...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s007260070076

    authors: Kanda-Nambu K,Yasuda Y,Tochikubo K

    更新日期:2000-01-01 00:00:00

  • The facile HPLC enantioresolution of amino acids, peptides on naphthylethylcarbamate-beta-cyclodextrin bonded phases using the acetonitrile-based mobile phase after their pre-column derivatization with phenyl isothiocyanate: factors that affect the resolu

    abstract::A variety of alpha-amino acids are enantioresolved for the first time on naphthylethylcarbamate-beta-cyclodextrin bonded phases (i.e., SN- and RN-beta-CD) using the acetonitrile-based mobile phase after their pre-column derivatization with phenyl isothiocyanate in alkaline medium. The resolution is better obtained on ...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-003-0038-4

    authors: Chen S

    更新日期:2004-06-01 00:00:00

  • Adenosine receptor agonists affect taurine release from mouse brain stem slices in ischemia.

    abstract::The release of the inhibitory amino acid taurine is markedly enhanced under ischemic conditions in both adult and developing brain stem, together with a pronounced increase in the release of the neuromodulator adenosine. We now studied the effects of adenosine receptor agonists and antagonists on [(3)H]taurine release...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-009-0351-7

    authors: Saransaari P,Oja SS

    更新日期:2010-05-01 00:00:00

  • A fusion antitumor peptide regulates proliferation and apoptosis of endothelial cells.

    abstract::The present research has been carried out to elicit the mechanism of antiangiogenic activity of a fusion peptide P2. Peptide P2 was designed by the connection of a heptapeptide MMP inhibitor to ES-2, a fragment of Endostatin. In a previous study, P2 demonstrated strong antiangiogenic and antitumor effect, and the curr...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-018-2589-4

    authors: Xu Y,Qiang X,Xing L,Wang H,Zhang J,Zhang F,Caliskan B,Wang M,Qiu Z

    更新日期:2018-08-01 00:00:00

  • Homocysteine and oxidative stress.

    abstract::Hyperhomocysteinemia is an independent risk factor for cardiovascular disease (ischemic disease, such as stroke and myocardial infarction, and arterial and venous thrombotic events) in the general population. We can assume that the association is causal, based on the example of homocystinuria, and on the evidence put ...

    journal_title:Amino acids

    pub_type: 杂志文章,评审

    doi:10.1007/s00726-003-0026-8

    authors: Perna AF,Ingrosso D,De Santo NG

    更新日期:2003-12-01 00:00:00

  • A comparative SAR study of thrombin receptor derived non peptide mimetics: importance of phenyl/guanidino proximity for activity.

    abstract::Thrombin, the most potent physiological platelet agonist interacts with cells through a specific G protein-coupled receptor which has been cloned and sequenced. Synthetic thrombin receptor peptides (TRAPs) comprising the first 5 amino acids (SFLLR and SFLLR-NH2) of the new N-terminus tethered ligand of the thrombin re...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/BF01318860

    authors: Alexopoulos K,Matsoukas J,Tselios T,Roumelioti P,Mavromoustakos T,Holada K

    更新日期:1998-01-01 00:00:00

  • The non-protein amino acid β-N-methylamino-L-alanine in Portuguese cyanobacterial isolates.

    abstract::The tailor made amino acid β-N-methyl-amino-L-alanine (BMAA) is a neurotoxin produced by cyanobacteria. It has been associated with certain forms of progressive neurodegenerative disease, including sporadic Amyotrophic Lateral Sclerosis and Alzheimer's disease. Some different reports of BMAA in cyanobacterial blooms f...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-011-1057-1

    authors: Cervantes Cianca RC,Baptista MS,Lopes VR,Vasconcelos VM

    更新日期:2012-06-01 00:00:00

  • The construction of an amino acid network for understanding protein structure and function.

    abstract::Amino acid networks (AANs) are undirected networks consisting of amino acid residues and their interactions in three-dimensional protein structures. The analysis of AANs provides novel insight into protein science, and several common amino acid network properties have revealed diverse classes of proteins. In this revi...

    journal_title:Amino acids

    pub_type: 杂志文章,评审

    doi:10.1007/s00726-014-1710-6

    authors: Yan W,Zhou J,Sun M,Chen J,Hu G,Shen B

    更新日期:2014-06-01 00:00:00

  • A convenient route to optically pure α-alkyl-β-(sec-amino)alanines.

    abstract::The cyclization of N-Boc-α-alkylserines to corresponding β-lactones under Mitsunobu reaction conditions and the ring opening with heterocyclic amines (pyrrolidine, piperidine, morpholine and thiomorpholine) produced N-Boc-α-alkyl-β-(sec-amino)alanines. The removal of the Boc group gives di-hydrochlorides of non-protei...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-011-1055-3

    authors: Olma A,Lasota A,Kudaj A

    更新日期:2012-06-01 00:00:00

  • Exploring pitfalls of 64Cu-labeled EGFR-targeting peptide GE11 as a potential PET tracer.

    abstract::The epidermal growth factor receptor (EGFR) represents an important molecular target for both radiotracer-based diagnostic imaging and radionuclide therapy of various cancer entities. For the delivery of radionuclides to the tumor, peptides hold great potential as a transport vehicle. With respect to EGFR, the peptide...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-018-2616-5

    authors: Striese F,Sihver W,Gao F,Bergmann R,Walther M,Pietzsch J,Steinbach J,Pietzsch HJ

    更新日期:2018-10-01 00:00:00

  • Microglial self-defence mediated through GLT-1 and glutathione.

    abstract::Glutamate is stored in synaptic vesicles in presynaptic neurons. It is released into the synaptic cleft to provide signalling to postsynaptic neurons. Normally, the astroglial glutamate transporters GLT-1 and GLAST take up glutamate to mediate a high signal-to-noise ratio in the synaptic signalling, and also to preven...

    journal_title:Amino acids

    pub_type: 杂志文章,评审

    doi:10.1007/s00726-011-0865-7

    authors: Persson M,Rönnbäck L

    更新日期:2012-01-01 00:00:00

  • The effects of red bull energy drink on human performance and mood.

    abstract::The effects of Red Bull Energy Drink, which includes taurine, glucuronolactone, and caffeine amongst the ingredients, were examined over 3 studies in a total of 36 volunteers. Assessments included psychomotor performance (reaction time, concentration, memory), subjective alertness and physical endurance. When compared...

    journal_title:Amino acids

    pub_type: 临床试验,杂志文章,随机对照试验

    doi:10.1007/s007260170021

    authors: Alford C,Cox H,Wescott R

    更新日期:2001-01-01 00:00:00

  • Possible sources and functions of L-homoarginine in the brain: review of the literature and own findings.

    abstract::L-Homoarginine is a cationic amino acid derivative, which is structurally related to L-arginine and lysine. Several lines of evidence point to nervous tissue as an important target of homoarginine action. In the mammalian brain homoarginine can be detected in noticeable quantities, but its origin is currently poorly e...

    journal_title:Amino acids

    pub_type: 杂志文章,评审

    doi:10.1007/s00726-015-1960-y

    authors: Bernstein HG,Jäger K,Dobrowolny H,Steiner J,Keilhoff G,Bogerts B,Laube G

    更新日期:2015-09-01 00:00:00

  • Abomasal infusion of arginine stimulates SCD and C/EBPß gene expression, and decreases CPT1ß gene expression in bovine adipose tissue independent of conjugated linoleic acid.

    abstract::Based on previous research with bovine peadipocytes, we hypothesized that infusion of arginine into the abomasum of Angus steers stimulates stearoyl-CoA desaturase (SCD) gene expression in bovine subcutaneous (s.c.) adipose tissue, and that this would be attenuated by conjugated linoleic acid (CLA). Growing Angus stee...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-013-1622-x

    authors: Choi SH,Wickersham TA,Wu G,Gilmore LA,Edwards HD,Park SK,Kim KH,Smith SB

    更新日期:2014-02-01 00:00:00

  • Mitochondrial nitric oxide inhibits ATP synthesis. Effect of free calcium in rat heart.

    abstract::Nitric oxide is a small potentially toxic molecule and a diatomic free radical. We report the interaction of L-arginine, oxygen and calcium with the synthesis of nitric oxide in heart mitochondria. Nitric oxide synthesis is increased in broken rat heart mitochondria compared with intact and permeabilized mitochondria....

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-002-0331-7

    authors: Saavedra-Molina A,Ramírez-Emiliano J,Clemente-Guerrero M,Pérez-Vázquez V,Aguilera-Aguirre L,González-Hernández JC

    更新日期:2003-01-01 00:00:00

  • Insilico study of the A(2A)R-D (2)R kinetics and interfacial contact surface for heteromerization.

    abstract::G-protein-coupled receptors (GPCRs) are cell surface receptors. The dynamic property of receptor-receptor interactions in GPCRs modulates the kinetics of G-protein signaling and stability. In the present work, the structural and dynamic study of A(2A)R-D(2)R interactions was carried to acquire the understanding of the...

    journal_title:Amino acids

    pub_type: 杂志文章

    doi:10.1007/s00726-012-1218-x

    authors: Prakash A,Luthra PM

    更新日期:2012-10-01 00:00:00