排序方式: 共有25条查询结果,搜索用时 15 毫秒
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Liu C Eriste E Sutton S Chen J Roland B Kuei C Farmer N Jörnvall H Sillard R Lovenberg TW 《The Journal of biological chemistry》2003,278(50):50754-50764
GPCR135, publicly known as somatostatin- and angiotensin-like peptide receptor, is expressed in the central nervous system and its cognate ligand(s) has not been identified. We have found that both rat and porcine brain extracts stimulated 35S-labeled guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) incorporation in cells over-expressing GPCR135. Multiple rounds of extraction, purification, followed by N-terminal sequence analysis of the ligand from porcine brain revealed that the ligand is a product of the recently identified gene, relaxin-3 (aka insulin-7 or INSL7). Recombinant human relaxin-3 potently stimulates GTPgammaS binding and inhibits cAMP accumulation in GPCR135 overexpressing cells with EC50 values of 0.25 and 0.35 nM, respectively. 125I-Relaxin-3 binds GPCR135 at high affinity with a Kd value of 0.31 nM. Relaxin-3 is the only member of the insulin/relaxin superfamily that can activate GPCR135. In situ hybridization showed that relaxin-3 mRNA is predominantly expressed in the dorsomedial ventral tegmental nucleus of the brainstem (aka nucleus incertus), as well as in discrete cells in the lateral periaqueductal gray and in the central gray nucleus. GPCR135 is expressed abundantly in the hypothalamus with discrete expression in the paraventricular nucleus of the hypothalamus and supraoptic nucleus, as well as in the cortex, septal nucleus, and preoptical area. Relaxin-3 has previously been shown to bind and activate the LGR7 relaxin receptor. However, we believe that neuroanatomical colocalization of GPCR135 and relaxin-3, coupled with a clear high affinity interaction, suggest that GPCR135 is the receptor for relaxin-3. The identification of relaxin-3 as the ligand for GPCR135 provides the framework for the discovery of a new brainstem/hypothalamus circuitry. 相似文献
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Kent Langel Staffan Lindberg Dana Copolovici Piret Arukuusk Rannar Sillard Űlo Langel 《International journal of peptide research and therapeutics》2010,16(4):247-255
Cell-penetrating peptides (CPPs) are able to efficiently internalize into cells and can therefore be used as vectors for non-viral
cellular delivery of different cargoes. Previous studies have shown that hydrophobic modifications of different CPPs can increase
their transfection efficiency dramatically. In this study we have modified the cell penetrating-peptide transportan 10 (TP10)
with a variety of hydrophobic molecules to determine the role of hydrophobicity in the uptake of these molecules. The results
can be used to synthesize more efficient delivery vectors. To evaluate how these constructs are able to transport cargoes
into cells we used 2′-OMe splice correcting oligonucleotides. Non-covalent peptide-cargo complexes were formed and their transfection
efficiency was measured using a luciferase readout system. The hydrophobicity of the novel modifications was correlated with
their biological efficacy. We determined the most efficient range of hydrophobicity for TP10 analogs for delivering oligonucleotides
into cells. In order to assess how the transfection efficacy of these particles is dependent on their size the hydrodynamic
diameter of the formed nanoparticles was measured using dynamic light scattering. These findings will be used to develop highly
efficient non-viral gene therapy vectors. 相似文献
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Markus Schueler Daniela?A. Braun Gayathri Chandrasekar Heon?Yung Gee Timothy?D. Klasson Jan Halbritter Andrea Bieder Jonathan?D. Porath Rannar Airik Weibin Zhou Joseph?J. LoTurco Alicia Che Edgar?A. Otto Detlef B?ckenhauer Neil?J. Sebire Tomas Honzik Peter?C. Harris Sarah?J. Koon Meral Gunay-Aygun Sophie Saunier Klaus Zerres Nadina?Ortiz Bruechle Joost?P.H. Drenth Laurence Pelletier Isabel Tapia-Páez Richard?P. Lifton Rachel?H. Giles Juha Kere Friedhelm Hildebrandt 《American journal of human genetics》2015,96(1):81-92
Nephronophthisis-related ciliopathies (NPHP-RC) are recessive diseases characterized by renal dysplasia or degeneration. We here identify mutations of DCDC2 as causing a renal-hepatic ciliopathy. DCDC2 localizes to the ciliary axoneme and to mitotic spindle fibers in a cell-cycle-dependent manner. Knockdown of Dcdc2 in IMCD3 cells disrupts ciliogenesis, which is rescued by wild-type (WT) human DCDC2, but not by constructs that reflect human mutations. We show that DCDC2 interacts with DVL and DCDC2 overexpression inhibits β-catenin-dependent Wnt signaling in an effect additive to Wnt inhibitors. Mutations detected in human NPHP-RC lack these effects. A Wnt inhibitor likewise restores ciliogenesis in 3D IMCD3 cultures, emphasizing the importance of Wnt signaling for renal tubulogenesis. Knockdown of dcdc2 in zebrafish recapitulates NPHP-RC phenotypes, including renal cysts and hydrocephalus, which is rescued by a Wnt inhibitor and by WT, but not by mutant, DCDC2. We thus demonstrate a central role of Wnt signaling in the pathogenesis of NPHP-RC, suggesting an avenue for potential treatment of NPHP-RC. 相似文献
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Liu C Kuei C Sutton S Chen J Bonaventure P Wu J Nepomuceno D Kamme F Tran DT Zhu J Wilkinson T Bathgate R Eriste E Sillard R Lovenberg TW 《The Journal of biological chemistry》2005,280(1):292-300
Insulin-like peptide 5 (INSL5) is a peptide that belongs to the relaxin/insulin family, and its receptor has not been identified. In this report, we demonstrate that INSL5 is a specific agonist for GPCR142. Human INSL5 displaces the binding of (125)I-relaxin-3 to GPCR142 with a high affinity (K(i) = 1.5 nM). In a saturation binding assay, (125)I-INSL5 binds GPCR142 with a K(d) value of 2.5 nM. In functional guanosine (gamma-thio)-triphosphate binding and cAMP accumulation assays, INSL5 potently activates GPCR142 with EC(50) values of 1.3 and 1.2 nM, respectively. In addition, INSL5 stimulates Ca(2+) mobilization in HEK293 cells expressing GPCR142 and G alpha(16). Overall, INSL5 behaves as an agonist for GPCR142 similar to relaxin-3. However, unlike relaxin-3, which is also a potent agonist for GPCR135 and LGR7, INSL5 does not activate either GPCR135 or LGR7. INSL5 inhibits (125)I-relaxin-3 binding to GPCR135 with a low potency (K(i) = 500 nM). A functional assay shows that INSL5 (1 microm) is a weak antagonist for GPCR135. In addition, INSL5 (up to 1 microm) shows no affinity or activity at LGR7 or LGR8 either in a binding assay or a bio-functional assay. Previously, we have demonstrated that GPCR142 mRNA is expressed in peripheral tissues, particularly in the colon. Here we show that INSL5 mRNA is expressed in many peripheral tissues, similar to GPCR142. The high affinity interaction between INSL5 and GPCR142 coupled with their co-evolution and partially overlapping tissue expression patterns strongly suggest that INSL5 is an endogenous ligand for GPCR142. 相似文献
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Eriste E Norberg A Nepomuceno D Kuei C Kamme F Tran DT Strupat K Jörnvall H Liu C Lovenberg TW Sillard R 《The Journal of biological chemistry》2005,280(42):35089-35097
A novel bioactive form of neurotensin post-translationally modified at a Glu residue was isolated from porcine intestine. Purification of the peptide was guided by detection of intracellular Ca2+ release in SK-N-SH neuroblastoma cells. Using high resolution accurate mass analysis on an ion trap Fourier transform mass spectrometer, the post-translational modification was identified as arginine linked to the gamma-carboxyl of Glu via an isopeptide bond, and we named the newly identified peptide "arginylated neurotensin" (R-NT, N-(neurotensin-C5-4-yl)arginine). Although arginylation is a known modification of N-terminal amino groups in proteins, its presence at a Glu side chain is unique. The finding places neurotensin among the few physiologically active peptides that occur both in post-translationally modified and unmodified forms. Pharmacologically, we characterized R-NT for its ligand activity on three known neurotensin receptors, NTR1, -2, and -3, and found that R-NT has similar pharmacological properties to those of neurotensin, however, with a slightly higher affinity to all three receptors. We expressed the intracellular receptor NTR3 as a soluble protein secreted into the cell culture medium, which allowed characterization of its R-NT and neurotensin binding properties. The creation of soluble NTR3 also provides a potential tool for neutralizing neurotensin action in vivo and in vitro. We have shown that SK-N-SH neuroblastoma cells express NTR1 and NTR3 but not NTR2, suggesting that the Ca2+ mobilization elicited by R-NT is via NTR1. 相似文献
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Janssen S Ramaswami G Davis EE Hurd T Airik R Kasanuki JM Van Der Kraak L Allen SJ Beales PL Katsanis N Otto EA Hildebrandt F 《Human genetics》2011,129(1):79-90
Bardet-Biedl syndrome (BBS) is a rare, primarily autosomal-recessive ciliopathy. The phenotype of this pleiotropic disease includes retinitis pigmentosa, postaxial polydactyly, truncal obesity, learning disabilities, hypogonadism and renal anomalies, among others. To date, mutations in 15 genes (BBS1-BBS14, SDCCAG8) have been described to cause BBS. The broad genetic locus heterogeneity renders mutation screening time-consuming and expensive. We applied a strategy of DNA pooling and subsequent massively parallel resequencing (MPR) to screen individuals affected with BBS from 105 families for mutations in 12 known BBS genes. DNA was pooled in 5 pools of 21 individuals each. All 132 coding exons of BBS1-BBS12 were amplified by conventional PCR. Subsequent MPR was performed on an Illumina Genome Analyzer II? platform. Following mutation identification, the mutation carrier was assigned by CEL I endonuclease heteroduplex screening and confirmed by Sanger sequencing. In 29 out of 105 individuals (28%), both mutated alleles were identified in 10 different BBS genes. A total of 35 different disease-causing mutations were confirmed, of which 18 mutations were novel. In 12 additional families, a total of 12 different single heterozygous changes of uncertain pathogenicity were found. Thus, DNA pooling combined with MPR offers a valuable strategy for mutation analysis of large patient cohorts, especially in genetically heterogeneous diseases such as BBS. 相似文献
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M Chaki R Airik AK Ghosh RH Giles R Chen GG Slaats H Wang TW Hurd W Zhou A Cluckey HY Gee G Ramaswami CJ Hong BA Hamilton I Cervenka RS Ganji V Bryja HH Arts J van Reeuwijk MM Oud SJ Letteboer R Roepman H Husson O Ibraghimov-Beskrovnaya T Yasunaga G Walz L Eley JA Sayer B Schermer MC Liebau T Benzing S Le Corre I Drummond S Janssen SJ Allen S Natarajan JF O'Toole M Attanasio S Saunier C Antignac RK Koenekoop H Ren I Lopez A Nayir C Stoetzel H Dollfus R Massoudi JG Gleeson SP Andreoli DG Doherty 《Cell》2012,150(3):533-548
Nephronophthisis-related ciliopathies (NPHP-RC) are degenerative recessive diseases that affect kidney, retina, and brain. Genetic defects in NPHP gene products that localize to cilia and centrosomes defined them as "ciliopathies." However, disease mechanisms remain poorly understood. Here, we identify by whole-exome resequencing, mutations of MRE11, ZNF423, and CEP164 as causing NPHP-RC. All three genes function within the DNA damage response (DDR) pathway. We demonstrate that, upon induced DNA damage, the NPHP-RC proteins ZNF423, CEP164, and NPHP10 colocalize to nuclear foci positive for TIP60, known to activate ATM at sites?of DNA damage. We show that knockdown of CEP164 or ZNF423 causes sensitivity to DNA damaging agents and that cep164 knockdown in zebrafish results in dysregulated DDR and an NPHP-RC phenotype. Our findings link degenerative diseases of the kidney and retina, disorders of increasing prevalence, to mechanisms of DDR. PAPERFLICK: 相似文献
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Andaloussi SE Lehto T Mäger I Rosenthal-Aizman K Oprea II Simonson OE Sork H Ezzat K Copolovici DM Kurrikoff K Viola JR Zaghloul EM Sillard R Johansson HJ Said Hassane F Guterstam P Suhorutšenko J Moreno PM Oskolkov N Hälldin J Tedebark U Metspalu A Lebleu B Lehtiö J Smith CI Langel U 《Nucleic acids research》2011,39(9):3972-3987