Structure–function relationship of different domains of the rat corticotropin-releasing factor receptor

Sydow, Sabine; Radulovic, Jelena; Dautzenberg, Frank M.; Spiess, Joachim

doi:10.1016/s0169-328x(97)00256-8

Cited by 36 publications

(53 citation statements)

References 44 publications

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“…This has been suggested, for example, for the interaction of a soluble portion of CCR5 with gp 120 (47), for which the affinity is reduced by a factor of 10 3 -10 4 . Consistent with our data, another study of a soluble ECD-1 of CRFR1 expressed in E. coli reported only low affinity binding to ovine CRF (43). The fact that both sauvagine and r/hCRF do not compete with astressin for binding to the bNT-CRFR1 may be due to the fact that binding of these two agonists requires coupling of the receptor to G-proteins.…”

Section: Discussionsupporting

confidence: 89%

“…Using mutational analyses, residues that affect binding of peptide or non-peptide antagonists as well as the selectivity of various CRF agonists have been identified in the extracellular and transmembrane domains of CRF receptors (22)(23)(24)(25)(41)(42)(43). All of the extracellular domains contribute, in some degree, to the binding of the CRF peptide ligands.…”

Section: Discussionmentioning

confidence: 99%

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Expression, Purification, and Characterization of a Soluble Form of the First Extracellular Domain of the Human Type 1 Corticotropin Releasing Factor Receptor

Perrin¹,

Fischer²,

Kunitake³

et al. 2001

Journal of Biological Chemistry

108

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Corticotropin releasing factor (CRF), 1 a 41-amino acid peptide, was identified initially on the basis of its primary role in the activation of the hypothalamic-pituitary-adrenal in response to stress. The CRF family of ligands includes sauvagine from frog and urotensin from fish as well as the additional mammalian family members, urocortin (1, 2), urocortin II (3, 4), and urocortin III (4, 5). Broader roles for CRF and its ligand family now involve effects on the cardiovascular, reproductive, gastrointestinal, immune, and central nervous systems (6 -8).The action of CRF and related ligands is initiated by binding to their receptors, which transduce an increase in intracellular cAMP. Thus far, two receptors, CRFR1 and CRFR2, have been cloned in mammals (9 -15). Homologous receptors have been identified in chicken (16), fish (17), and Xenopus (18) and a third receptor, CRFR3, with high levels of sequence identity to CRFR1, has recently been cloned in catfish (17). Both CRFR1 and CRFR2 exist as multiple splice variants and belong to the type B 7-transmembrane receptor family that includes receptors for growth hormone releasing factor, secretin, calcitonin, vasoactive intestinal peptide, glucagon, glucagon-like peptide, and parathyroid hormone (PTH). Receptors for the CRF ligand family have been characterized in the central nervous system, pituitary, gastrointestinal tract, epididymis, heart, gonads, and adrenals (6).The affinities of CRF and urocortin in binding to CRFR1 are nearly the same but in binding to CRFR2, urocortin is ϳ10 times more potent than CRF (19). Both urocortins II and III are highly selective in binding and activating CRFR2 compared with CRFR1 (4, 5). A synthetic peptide antagonist, astressin, binds with equally high affinity to CRFR1 and CRFR2 (19,20).The CRF receptor family consists of proteins with a relatively large first extracellular domain (ECD-1). Mutagenesis studies have identified regions of the CRF receptors that are implicated in differential recognition of agonists and antagonists, as well as in governing the ligand selectivity of the two types of receptors (21-25). We showed that a chimeric receptor in which the ECD-1 of CRFR1 replaced the ECD of the activin receptor, a single transmembrane receptor kinase (26), was capable of high affinity binding to both astressin and urocortin (21). The mode of receptor activation was explored by our study of a tethered peptide-receptor chimera in which the first 16 amino acids of CRF were substituted for the ECD-1 of CRFR1 (CRF (1-16)/R1 ⌬N ) (27). This chimera displayed continual signaling suggesting that the N-terminal third of CRF, when presented in proximity to the receptor, is able to cause activation.Biochemical characterizations of soluble ECD-1s include those of receptors for follicle stimulating hormone (28), luteinizing hormone/human chorionic gonadotropin (29), calcium (30), PTH (31), glucagon-like peptide-1 (32), and glutamate (33). The ECD-1 of the metabotropic glutamate receptor has

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Expression, Purification, and Characterization of a Soluble Form of the First Extracellular Domain of the Human Type 1 Corticotropin Releasing Factor Receptor

Perrin¹,

Fischer²,

Kunitake³

et al. 2001

Journal of Biological Chemistry

108

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“…Thus, the species of 52, 60 and 70 kDa likely represent glycosylated forms of CRF1 (α and/or c), while the 80-85 kDa protein might represent a dimer, in agreement with reports of varying CRF1 mobility on SDS-PAGE (Grigoriadis, De Souza, 1988;Grigoriadis, De Souza, 1989;Ruhmann et al, 1996;Hauger et al, 2000;Slominski et al, 2006a;Slominski et al, 2006b). In fact, glycosylated forms of CRF1 of 70 kDa and 53 kDa were respectively detected in pituitary and brain tissues and skin cells (Grigoriadis, De Souza, 1988;Grigoriadis, De Souza, 1989;Slominski et al, 2006a), whereas multiple CRF1 bands were detected in the AtT20 and Y-79 cell lines, and in rat pituitary and human skin cells (Sydow et al, 1997;Slominski et al, 2006a;Slominski et al, 2006b). As regards bands of lower MW (37 and 40 kDa), these could represent products of proteolytic processing of CRF1.…”

Section: Detection Of Crf1 Proteins In Cos-7 Cellssupporting

confidence: 87%

Molecular cloning and initial characterization of African green monkey (Cercopithecus aethiops) corticotropin releasing factor receptor type 1 (CRF1) from COS-7 cells

Slominski

Żmijewski

Pisarchik

et al. 2007

Gene

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We report the expression of endogenous CRF1 in COS-7 cells (African green monkey origin). Cloning of the coding region of CRF1 gene identified three alternatively spliced isoforms with nucleotide and predicted amino acid sequences corresponding to the membrane bound α and c and soluble e isoforms. DNA sequencing of the main isoform CRF1α showed homologies of 99%, 97% and 91% with the rhesus monkey, human and rodent genes, respectively; the deduced protein sequence differed in only one amino acid with Rhesus monkey and human. Western blot analysis with antibodies against human CRF1 demonstrated immunoreactive proteins with MW of 37, 52, 70 and 80-85 in crude membrane or cytoplasm preparation; two additional species of 40 and 60 kDa were detected only in the cytoplasmic fraction. On immunocytochemistry CRF1 was localized to both the cell surface and intracellularly. The receptor was functional, e.g., addition of CRF to COS-7 cells inhibited cell proliferation and stimulated release of arachidonic acid; nevertheless, it was poorly coupled to cAMP production (its stimulation was minimal in native cells). In conclusion, COS cells that are routinely used for the study of transfected CRF receptors do express endogenous CRF1 mRNA with splicing behavior similar to that reported in human and rodent cells, and translated into functional CRF1 receptors.

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“…It was demonstrated by CD that ␣-helical structures of h͞rCRF and ␣-hel-CRF 9-41 are involved in binding to CRFBP (32). In view of the crucial role of Ala 22 for binding to CRFBP, we concluded that the hydrophobic patch may be important for binding to CRFBP. Consistently, Ala 24 located on The relative hydrophobicities were taken from the consensus hydrophobicity scale of Eisenberg (24).…”

Section: Discussionmentioning

confidence: 77%

“…This interaction may have been disturbed by the charged bulky Glu residue in [Glu 22 ]h͞ rCRF as indicated by its lower affinity, whereas it may have been facilitated in [Ala 21 ]Svg as demonstrated by the latter peptide's high affinity to CRFBP. This view is consistent with the observation that replacement of Thr 22 of oCRF by an Ala residue enhanced the affinity by one order of magnitude (12).…”

Section: Discussionmentioning

confidence: 99%

A single amino acid serves as an affinity switch between the receptor and the binding protein of corticotropin-releasing factor: Implications for the design of agonists and antagonists

Eckart

Jahn

Radulovic

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

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In view of the observation that corticotropin-releasing factor (CRF) affects several brain functions through at least two subtypes of G protein-dependent receptors and a binding protein (CRFBP), we have developed synthetic strategies to provide enhanced binding specificity. Human͞rat CRF (h͞rCRF) and the CRF-like peptide sauvagine (Svg), differing in their affinities to CRFBP by two orders of magnitude, were used to identify the residues determining binding to CRFBP. By amino acid exchanges, it was found that Ala 22 of h͞rCRF was responsible for this peptide's high affinity to CRFBP, whereas Glu 21 located in the equivalent position of Svg prevented high affinity binding to CRFBP. Accordingly, [Glu 22 ]h͞rCRF was not bound with high affinity to CRFBP in contrast to [Ala 21 ]Svg, which exhibited such high affinity. Furthermore, the affinity of both peptides to either CRF receptor (CRFR) subtype was not reduced by these replacements, and their subtype preference was not changed. Thus, exchange of Ala and Glu and vice versa in positions 22 and 21 of h͞rCRF and Svg, respectively, serves as a switch discriminating between CRFBP and CRFR. On the basis of this switch function, development of new specific CRF agonists and antagonists is expected to be facilitated. One application was the modification of the CRF antagonist astressin (Ast), whose employment in animal experiments is limited by its low solubility in cerebrospinal fluid. Introduction of Glu residues into Ast generated with [Glu 11,16 ]Ast an acidic astressin, which efficiently antagonized in vivo the CRFR1-dependent reduction of locomotion induced by ovine CRF without detectable binding to CRFBP.

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Structure–function relationship of different domains of the rat corticotropin-releasing factor receptor

Cited by 36 publications

References 44 publications

Expression, Purification, and Characterization of a Soluble Form of the First Extracellular Domain of the Human Type 1 Corticotropin Releasing Factor Receptor

Expression, Purification, and Characterization of a Soluble Form of the First Extracellular Domain of the Human Type 1 Corticotropin Releasing Factor Receptor

Molecular cloning and initial characterization of African green monkey (Cercopithecus aethiops) corticotropin releasing factor receptor type 1 (CRF1) from COS-7 cells

A single amino acid serves as an affinity switch between the receptor and the binding protein of corticotropin-releasing factor: Implications for the design of agonists and antagonists

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