MXPA00012127A - Novel angiotensin receptor, production and use thereof - Google Patents

Abstract

The invention relates to a novel angiotensin receptor and to the production and use thereof. The novel angiotensin receptor hat at least one subunit containing the amino acid sequence SEQ ID NO. 10.

Description

NOVEDOUS ANGIOTENSIN RECEPTOR, ITS PRODUCTION AND USE The invention relates to a novel receptor for angiotensin, its preparation and its use. The renin-angiotensin system plays an important role in the regulation of the cardiovascular system and L electrolyte and fluid balance. Physiologically active effector hormones, for example, angiotensin IC (Angll) and angiotensin 1-7 (Ang-7) are produced by proteolytic dissociation of the precursor angiotensinogen that is synthesized in the liver, and, respectively, angiotensin I, which is formed from angiotensinogen A large number of proteases are involved in these dissociations (eg, renin and angiotensin conversion enzyme (ACE)). To date, two human angiotensin receptors have been described that have been designated ATI and AT2. Both receptors are angiotensin II (Ang II) receptors, that is, these receptors bind in preference to Ang II. The cDNA sequences of these two receptors (Takayanagi, R. et al (1992), Biochemical and Biophysical Research Communications 183, 910-915; Tsuzuki, S. Et al. (1994). Biochemical and Biophysical Research Communications 200, 1449. -1454) each possess seven hydrophobic regions that constitute domains of. potential transmembrane. Due to this characteristic domain structure, it is considered that the receivers of He? Angiotensin ATI and AT2 belong to the family of transmembrane receptors 7 that mediate the effect of angiotensin II ligand through signaling transduction cascades coupled with intracellular protein G. Due to the pharmacological effects mediated by other angiotensins that differ from Angll and / or from the peptides (fragments formed endogenously of Angi and Anglli derived from angiotensin I and angiotensin II, respectively, it has been suspected that there must be other human angiotensin receptors In connection with this aspect, a receptor for the heptapeptide Angl-7 has recently been requested (Ferrario, CM et al (1997) Hypertension 30, 535-541) The present invention relates to a novel receptor of angiotensin having at least one subunit containing the amino acid sequence SEQ ID NO: 10, with this amino acid sequence possibly possessing additional amino acids at the C-terminus The angiotensin receptor is a transmembrane receptor. with the previously described angiotensin1 receptors, it preferably has less than seven transmembrane domains.The subunit containing the SEQ sequence IEi NO: 10 preferably has a transmembrane domain. This transmembrane domain is preferably more than 6, preferably 8 or 9, especially 7 amino acids in length.

The transmembrane domain preferably has the sequence "MSIVIPL" (written with the unique letter code for amino acids). The angiotensin receptor may possess one or more subunits. For example, the receptor can be present either as a) a monomer (a subunit containing the sequence SEQ ID NO: 10), or as b) a dimer, which contains an identical subunit or a different subunit, or as c) a tetramer, which contains three additional identical subunits or which contains an identical subunit and two different subunits. However, the angiotensin receptor may also be present as a monomer (a subunit) or as a dimer (two different subunits) until the ligand has been bound and changed only in a dimeric or tetrameric form, respectively, as a result of ligand binding. Receptors that have a structure similar to the structure of the novel angiotensin receptor, are, among others, the insulin receptor, growth factors, such as for example nerve growth factor (NGF), endothelial growth factor d1 (EGF), factor of growth d 'fibroblast (FGF), platelet derived growth factor (PDGF), T cell antigen receptors, ^ ü ^ ßa ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ l ^^^^^ ^^^^^^^^^^^^ cytokine atopoietic receptors (eg IL1, IL5), and adhesion molecules (integrins and selectins). If appropriate, the novel angiotensin receptor can be modified after translation; for example, the receptor can be glycosylated and / or phosphorylated. The angiotensin receptor can bind its ligand (s), it can mediate the effect of this / these ligand (s) intracellularly. The angiotensin receptor can be linked to angiotensin 1-7 and / or an angiotensin derivative 1-7. Angiotensin derivatives 1-7 are preferably truncated peptides derived from Angl-7, for example, products of the degradation of Angl-7. Examples of derivatives of Ang-7 or angiotensin III and angiotensin IV. The receiver binds preference to Angl-7. The invention relates to a receptor that possesses or contains the amino acid sequence illustrated in Table 2, SEQ ID NO: 10. The angiotensin receptor or subunit containing the sequence SEQ ID NO: 10 has at least 175 amino acids of length. The invention further relates to a nucleic acid that poses a sequence encoding the novel angiotensin receptor ("receptor"). For example, the receptor can be encoded, in whole or in part, by a nucleic acid possessing the sequence SEQ ID NO: 9. A nucleic acid encoding the receptor or a subunit of the receptor can contain the sequence SEQ ID NO: 9 and, possibly, also possess nucleotides belonging to the coding sequence, at the 3 'end. The nucleic acid can be, for example, a DNA, a cDNA or an RNA. The nucleic acid can be a gene, it being possible for the sequence SEQ ID NO: 9 to be interrupted by non-coding sequences (introns). If appropriate, the nucleic acid can be derived. A nucleic acid derivative can be, for example, a salt or a nucleic acid derivative containing modified nucleotides in addition to natural nucleotides or consisting entirely of modified nucleotides. The invention also relates to a process for the preparation of a nucleic acid encoding the angiotensin receptor and containing the sequence SEQ ID NO: 9. For example, said nucleic acid can be amplified in a polymerase chain reaction that employs oligonucleotides as primers, whose sequences correspond to the homologous regions (conserved regions) in the nucleotide sequences of the angiotensin ATI and AT2 receptors and cDNA which is preferably prepared from a tissue (or cells) in which angiotensin 1-7, or a derivative thereof, has an effect and / or from a tissue (or cell) in which ATI or AT2 are not expressed. In a particular mode of the process, degenerate initiators are used whose sequences are derived from the homologous regions of the ATI and AT2 sequences. Preferably Ang3A and Ang3B primers are used for this purpose. The cDNA that has been prepared from endothelial cells can be used, for example, as a template. For example, the cDNA prepared from HUVECs (human umbilical vein endothelial cells) can be used as an anneal. The invention also relates to a process for the preparation of a novel receiver. For example, the novel angiotensin receptor can be prepared by integrating a nucleic acid whose sequence encodes the angiotensin receptor and preferably contains the sequence SEQ ID NO: 9, into a suitable vector, with introduction of the recombinant vector into a cell and with expression of the angiotensin receptor in this cell. The invention further relates to the use of a nucleic acid encoding the novel angiotensin receptor and preferably containing the sequence SEQ ID NO: 9, for example, in a process for the preparation of a recombinant angiotensin receptor, with the introduction of the nucleic acid that is under the control of a suitable promoter in a cell and its expression in this cell. The invention further relates to the use of a nucleic acid encoding the novel angiotensin receptor and which: preferably contains the sequence SEQ ID NO: 9, for the preparation of a recombinant cell in which the angiotensin receptor can be expressed. The invention relates to a recombinant cell which expresses an angiotensin receptor having the sequence SEQ ID NO: 10. The invention further relates to the use of a nucleic acid encoding the angiotensin receptor and preferably containing the sequence SEQ ID NO: 9 for the production of a drug for the treatment and prevention of diseases accompanied by defective regulation of the cardiovascular system, the electrolyte balance or L fluid balance. The invention relates to a drug comprising a nucleic acid having the sequence SEQ ID NO: 9. The invention further relates to the use of a nucleic acid encoding the angiotensin receptor and preferably containing the sequence SEQ ID NO: 9 for the production of a transgenic animal that does not contain the corresponding angiotensin receptor gene or for the production of a transgenic animal that overexpresses the corresponding angiotensin receptor gene. Transgenic animals can be used, for example, for the characterization of the specificity and biochemical function of the angiotensin receptor. The invention relates to a non-human transgenic animal that expresses the angiotensin receptor. The invention also relates to the use of a nucleic acid which encodes the angiotensin receptor and preferably contains the sequence SEQ ID NO: 9 in processes to identify and characterize substances that can be used as angiotensin receptor antagonists and agonists and / or in processes in which substances that inhibit or Activate the functional expression of the encoded angiotensin receptor are identified. The invention relates to a process for identifying and characterizing angiotensin receptor agonists or antagonists with a nucleic acid having the sequence SEQ ID NO: 9 introduced into a cell and the angiotensin receptor expressed in the cell, this cell is incubated with a substance to be investigated and the effect of this substance on the angiotensin receptor is determined. The invention also relates to the use of a nucleic acid encoding the angiotensin receptor and preferably containing the sequence SEQ ID NO: 9, in gene therapy or for the production of a drug that can be used in gene therapy. In addition to this, the invention relates to the use of a nucleic acid encoding the angiotensin receptor and preferably containing the (gap) SEQ ID NO: 9) as a tool in molecular biology (eg, as a probe). The invention also relates to the use of the novel angiotensin receptor. For example, the receiver of Angiotensin can be used in processes to identify and characterize substances that can be used as receptor agonists or antagonists (e.g., binding assays and functional assays for screening). In addition, the angiotensin receptor or parts thereof, eg, a predetermined epitope, for example, by the amino acid sequence SEQ ID NO: 10 or a portion thereof, can be used to prepare antibodies in accordance with known methods (Harlow and Lane, "Antibodies-A Laboratory Manual," Cold Spring Harbor Laboratory, ISBN-0--87969-314-2). 7 Antibodies that have been prepared in this way and that bind specifically to the novel angiotensin receptor can be used to detect the. novel receptor, for example, in diagnostic methods and / or to subject the recipient to further characterization. DETAILED DESCRIPTION OF THE INVENTION In order to find the novel receptor, the known sequences encoding the angiotesin II ATI and AT2 receptors were used to synthesize oligonucleotides whose sequences correspond to the conserved regions of the AT1 / AT2 coding sequences or which they focus on these conserved regions (Ang3A and Ang3B). These oligonucleotides were used in polymerase chain reactions using cDNA from human HUVECs (human umbilic vein endothelial cell) as an annealing. This l -m u ¿?átátá? aiamÉ > i <íi ^? My cell line is derived from endothelial cells that line the blood vessels. These cells do not express any ATI or AT2 receptor. A DNA fragment having an open reading frame and a nucleotide sequence (SEQ ID NO: 3) that had not been previously described was amplified and identified. This nucleotide sequence did not show any concordance with sequences in the Genbank® or EMBL databases. The comparison of this sequence with sequences in the base of Lifeseq® data using the "Blast" program modules and "Assembly" (GGC program packages) showed that SEQ ID NO: 3 had junction with clones Nos. 1458429, 3341232, 3440195, 1250818, 2836520, 1310286 and 1362205 of Lifeseq © (ESTs of unknown function). The theoretical DNA fragment of 0.5 kbp that could be derived from this virtual contiguous (3) was confirmed experimentally (example 2, polymerase chain reaction using the pair of primers Ang4A and Ang4B). The sequence of the DNA fragment, together with the open reading frame contained there, was elucidated (SEQ ID NO: 9) by extension of 5 '(example 3) in the sequence of' vector. The sequence SEQ ID NO: 9 also contains the region not translated 5 'or a part thereof. An analysis of the 5 'region of the nucleotide sequence of' SEQ ID NO: 9 showed that, upstream of the triplet of ' considered start methionine (ATG), a purine nucleotide at position -3 and cytosine nucleotides at positions -1 and -4 were located. The location of these nucleotides at positions -1, -3 -4 corresponds to the eukaryotic consensus sequence for the starting point for the initiation of an mRNA translation. The reading frame opened in SEQ ID NO: 9 encodes a protein containing at least 175 amino acids (SEQ ID NO: 10). A hydrophobicity analysis of this amino acid sequence, performed by the GCG program, showed that the sequence possesses a hydrophobic domain of approximately "amino acids in length at the N-terminus. This hydrophobic domain can represent a transmembrane domain that is employed. to anchor this novel receptor in the membrane.In contrast, the sequence possesses a domain at the C-terminus that is more or less hydrophilic.Examples: Example 1: The oligonucleotide primers Ang3A and Ang3B, which, based on the genes of ATI / 2 receptor, containing a DNA fragment of 0.19 kbp, were used for the first polymerase chain reaction, cDNA was used from a commercially available HUVEC cDNA library (Stratagene catalog No. 937223, Stratagene GmbH, Heidelberg) as templated DNA 25 Ang3A nucleotide sequence: ^^^^^^^^^^^^^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^^^ SEQ ID NO: 1: 5 '-TTG TKC TKK YCT TY TC TTT SCT GGM TTC CC- 3' Nucleotide sequence of Ang3B: SEQ ID NO: 2: 5 '-TTY CCM ASA AAR CMA TAM ARA ARM GGA TT-3 'where M = (A / C) Y = (C / T) R = (A / G) S = (G / C) K = (G / T) W = (A / T) ). For the polymerase chain reaction, 1 μg of cDNA was incubated from the HUVEC library with recombinant DNA polymerase Thermus aquaticus (Taq) (Boehringer Mannheim, Germany), and 10 pmol each of the Ang3A and Ang3B primers by 50 μl of reaction assay in a thermal cycler (Perkin Elmer 2400, thermal cycler, Perkin Elmer Applied Biosystems, Weiterstadt). The polymerase chain reaction was carried out using the following temperature profile: 20 seconds at 95 ° C, 10 seconds at 38 ° C and 20 seconds at 72 ° C (60 cycles). A DNA fragment of 130 base pairs (bp) in length was amplified. This DNA fragment of 130 base pairs was ligated into the pCR 2.1 TOPO® vector (Invitrogen BV, NV Leek, the Netherlands) and transformed into E. coli ToplOF '. The fragment sequence of 'DNA of 130 base pairs present in pCR2.1 was determined using a capillary electrophoresis sequencer (ABI 310, Applied Biosystems, eiterstadt) (SEQ ID NO: 3). Example 2: The sequence SEQ ID NO: 3 was compared with the sequences 5 contained in the publicly available databases (for example, EMBL and GenBank®) using the program modules FASTA and TFASTA (University of Wisconsin Genetics Computer Group of Genetic Computation) United States of America, programmatic package). In addition, the sequence was used for a comparison of corresponding homology with the sequences that are included in the Lifeseq® database of the company Incyte (Palo Alto, United States of America). Sequences of expressed sequence markers (ESTs) that are spliced with SEQ ID NO: 3 were identified in the Lifeseq® database (Lifeseq® clones Nos. 1458429, 3341232, 3440195, 1250818, 2836520, 1310286 and 1362205). The Ang4A and Ang4B primers were synthesized based on the sequence that the inventors assembled by the fact d ' order the expressed sequence markers that are spliced (ESTs). Nucleotide sequence of Ang4A: SEQ ID NO: 4: 5 '-TGG GGG TTG ATA CAG CAG AGA C-3' Nucleotide sequence of Ang4B: 25 SEQ ID NO: 5: 5 '-GCA CTG CCC TCT CTT TAT CCA AA -3' Polymerase chain reactions were carried out in each case with 0.1 μg of cDNA from the HUVEC library, as a template, and 10 pmol each of the Ang4A and Ang4B primers by 50 μl of the reaction assay and the polymerase mixture was used. of Advantage cDNA (Clontech, Heidelberg). The following temperature profile was used for the polymerase chain reactions: 30 seconds at 94 ° C and 2 minutes with 30 seconds at 60 ° C (60 cycles). A DNA fragment of 500 base pairs in length was obtained. This fragment was cloned and sequenced (SEQ ID NO: 6) according to that described in example 1). The sequence has an open reading frame (ORF). Example 3: Ang5A and Ang5B primers were synthesized to elucidate the 500 base pair DNA fragment sequence in the 5 'region. Ang5A nucleotide sequence: SEQ ID NO.7: 5 '-TGG GGA TTG ATA GGC AG-3' Nucleotide sequence of Ang5B: SED ID NO.8: 5 '-ATA ACT GTT GGA TTT CTC AA-3' The nucleotide sequence of Ang5A and Ang5B in each case corresponds to a part of the DNA fragment sequence of 500 base pairs. These two primers are used in additional polymerase chain reactions using the reverse M13 and direct M13 primers (-20), respectively.

* ^ L ^ ¡nj ^ g ^ jg ^ ¡^ Sg - ^^^^ j The reverse M13 and direct M13 primers (-20) correspond to the sequences of the bacteriophage Uni-ZAP® vector to prepare the cDNA library of HUVEC. These polymerase chain reactions were carried out employing in each case 0.1 μg of cDNA from the HUVEC library as annealed and 10 pmol each of the Ang5A or Ang5B primers and, respectively, reverse M13 or direct M13 (-20) (a from the cloning kit Topo-TA (Clontech)) and Taq polymerase (Boehringer Mannheim) and also Taq Start® antibodies (Clontech). The polymerase chain reactions were performed using the following temperature profile: 30 seconds at 96 ° C, 30 seconds at 50 ° C, and 45 seconds at 72 ° C (70 cycles). This resulted in obtaining a DNA fragment that has; a total length of 660 base pairs. This fragment was; cloned and sequenced according to that described in example 1. The DNA fragment has the sequence SEQ ID NO: 9. The sequence SEQ ID NO: 9 contains the region of; 5 'whole coding. The GCG modules assemble, map, move, structure; Graphing and stacking were also employed to perform the computer-assisted sequence analysis in addition to the programmatic modules that have already been mentioned. Table 1: SEQ ID NO. 9 aaaaj || I || t ta ¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡(partial) sequence of nucleotides of the angiotensin receptor 1 TCGTGTGTGA ACATCACAGG GTTTGTGGAT GCACTTAGAT GTTTGCAATG 51 AGCACTGTGG CTGGCATGCC CCAGTGTTTT GGATACCAAT GCATAGGACT 101 CCATAGTAAT CGAATTTACC AGAGGCGAAC GTCATGAGCA TAGTGATCCC 151 ATTGGGGGTT GATACAGCAG AGACGTCATA CTTGGAAATG GCTGCAGGTT 201 CAGAACCAGA ATCCGTAGAA GCTAGCCCTG TGGTAGTTGA GAAATCCAAC 251 AGTTATCCCC ACCAGTTATA TACCAGCAGC TCACATCATT CACACAGTTA 301 CATTGGTTTG CCCTATGCGG ACCATAATTA TGGTGCTCGT CCTCCTCCGA 351 CACCTCCGGC TTCCCCTCCT CCATCAGTCC TTATTAGCAA AAATGAAGTA 401 GGCATATTTA CCACTCCTAA TTTTGATGAA ACTTCCAGTG CTACTACAAT 451 CAGCACATCT GAGGATGGAA GTTATGGTAC TGATGTAACC AGGTGCATAT 501 GTGGTTTTAC ACATGATGAT GGATACATGA TCTGTTGTGA CAAATGCAGC 551 GTTTGGCAAC ATATTGACTG CATGGGGATT GATAGGCAGC ATATTCCTGA 601 TACATATCTA TGTGAACGTT GTCAGCCTAG GAATTTGGAT AAAGAGAGGG 651 CAGTGCTACT Table 2: SEQ ID NO. 10 (partial) amino acid sequence of the novel receptor 1 MSIVIPLGVDTAETSYLE AAGSEPESVEASPVWEKSNSYPHQLYTSSS 51 HHSHSYIGLPYADHNYGARPPTPTPPASPPPSVLISKNEVG IFTTPNFDET 101 SSATTISTSEDGSYGTDVTRCICGFTHDOGY ICCDKCSV WQHIDC GID 151 RQHIPDTYLCERCQPRNLDKERAVL Table 3: SEQ ID NO. 3 1 CTCCTCCATC AGTCCTTATT AGCAAAAATG AAGTAGGCAT ATTTACCACT 51 CCT ATTTTG ATGAAACTTC CAGTGCTACT ACAACCAGAG CGAG Table 4: SEQ ID NO: 1 June ATTGGGGGTT GATACAGCAG AGACGTCATA CTTGGAAATG GCTGCAGGTT 51 CAGAACCAGA ATCCGTAGAA GCTAGCCCTG TGGTAGTTGA GAAATCCAAC 101 AGTTATCCCC ACCAGTTATA TACCAGCAGC TCACATCATT CACACAGTTA 151 CATTGGTTTG CCCTATGCGG ACCATAATTA TGGTGCTCGT CCTCCTCCGA 201 CACCTCCGGC TTCCCCTCCT CCATCAGTCC TTATTAGCAA AAATGAAGTA 251 GGCATATTTA CCACTCCTAA TTTTGATGAA ACTTCCAGTG CTACTACAAT 301 CAGCACATCT GAGGATGGAA GTTATGGTAC TGATGTAACC AGGTGCATAT 351 GtGGTTTTAC ACATGATGAT GGATACATGA TCTGTTGTGA CAAATGCAGC 401 GTTTGGCAAC ATATTGACTG CATGGGGATT GATAGGCAGC ATATTCCTGA 451 TACATATCTA TGTGAACGTT GTCAGCCTAG GAATTTGGAT AAAGAGAGGG 501 CAGTGCTACT LIST OF SEQUENCES (1) GENERAL INFORMATION: (i) Applicant: (A) NAME: Hoechst Marion Roussel Deutschland GMBH (B) STREET: - (C) CITY: Frankfurt (D) FEDERAL STATE: - (E) COUNTRY: Germany ( F) POSTAL CODE: 65926 (G) TELEPHONE: 069-305-7072 (H) FAX: 069-35-7175 (I) TELEX: - (li) TITLE OF THE INVENTION: RECEIVER NOVEDOSO ANGIOTENSINA, ITS PRODUCTION AND USE ( iii) NUMBER OF SEQUENCES: 10 (iv) COMPUTER LEGIBLE FORMAT: (A) TYPE OF MEDIUM: soft disk (B) COMPUTER: compatible with IBM PC (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) PROGRAMMING: Patentln Relay # 1.0, Version tf 1.30 (EPO) (2) INFORMATION FOR SEQ ID NO: l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 32 base pairs (B) TYPE: nucleotide (C) NUMBER OF CHAINS: unique (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: genomic DNA (ix) CHARACTERISTICS: 5 (A) NAME / KEY: exon (B) LOCATION: 1..32 (D) INFORMATION MISCELLANEOUS: / note = "M = (A / C); Y (C / T); R = (A / G); S = (G / C); K = (G / T); W (A / T) ) "10 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1: TTGTKCTKKY CTTYWTCWTT TSCTGGMTTC CC 32 (2) INFORMATION FOR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: 15 (A) LENGTH: 29 base pairs (B) TYPE: nucleotide (C) NUMBER OF CHAINS: single (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: genomic DNA 20 (ix) CHARACTERISTICS: (A) NAME / KEY: exon (B) LOCATION: 1..29 (D) MISCELLANEOUS INFORMATION: / note = "M = (A / C); Y ( C / T), R = (A / G), S = (G / C), K = (G / T), W 25 (A / T) " »'- ~ .r. -. < - .. ~ - jé? H & A (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: TTYCCMASAA ARCMATAMAR AARMGGATT 29 (2) INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 94 base pairs (B) TYPE: nucleotide (C) NUMBER OF CHAINS: single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: genomic DNA (ix) CHARACTERISTICS: (A) NAME / KEY: exon (B) LOCATION: 1..94 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: CTCCTCCATC AGTCCTTATT AGCAAAAATG 30 AAGTAGGCAT ATTTACCACT CCTAATTTTG 60 ATGAAACTTC CAGTGCTACT ACAACCAGAG CGAC 94 (2) INFORMATION FOR SEQ ID NO:: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleotide (C) NUMBER OF CHAINS: Single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Genomic DNAKEY: exon (B) LOCATION: 1..22 (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 4: TGGGGGTTGA TACAGCAGAG AC 22 (2) INFORMATION FOR SEQ ID NO: 5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 base pairs (B) TYPE: nucleotide (C) NUMBER OF CHAINS: single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: genomic DNA (ix) CHARACTERISTICS: (A) NAME / KEY: exon (B) LOCATION: 1..23 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: GCACTGCCCT CTCTTTATCC AAA 23 (2) INFORMATION FOR SEQ ID NO: 6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 510 base pairs (B) TYPE: nucleotide (C) NUMBER OF CHAINS: single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: genomic DNA (ix) CHARACTERISTICS: (A) NAME / KEY: exon (B) LOCATION: 1..510 (xi) SEQUENCE DESCRITION: SEQ ID NO: 6: ATTGGGGGTT GATACAGCAG AGACGTCATA CTTGGAAATG GCTGCAGGTT CAGAACCAGA 60 ATCCGTAGAA GCTAGCCCTG TGsTAGTTGA GAAATCCAAC AGTTATCCCC ACCAGTTATA 120 TACCAGCAGC TCACATCATT CACACAGTTA CATTGGTTTG CCCTATGCGd ACCATAATTA 180 TGGTGCTCGT CCTCCTCCGA CACCTCCGGC TTCCC TCCT CCATCAGTCC TTATTAGCAA 240 AAATGAAGTA GGCATATTTA CCACTCCTAA TTTTGATGAA ACTTCCAGTG CTACTACAAT 300 CAGCACATCT GAGGATGGAA GTTATGGTAC TGATGTAACC AGGTGCATAT GTG3TTTTAC 360 ACATGATGAT GGATACATGA TCTGTTGTGA CAAATGCAGC GTTTGGCAAC ATATTGACTG 420 CATGGGGATT GATAGGCAGC ATATTCCTGA TACATATCTA TGTGAACGTT GTCAGCCTAG_480_GAATTTGGAT AAAGAGAGGG CAGTGCTACT 510 (2) INFORMATION FOR SEQ ID NO: 7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleotide (C) NUMBER OF CHAINS: single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: genomic DNA (ix) CHARACTERISTICS: (A) NAME / KEY: exon (B) LOCATION: 1..17 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7: ¿S * -. .s & ts TGGGGATTGA TAGGCAG 17 (2) INFORMATION FOR SEQ ID NO: 8: (i) SEQUENCE CHARACTERISTICS: 5 (A) LENGTH: 20 base pairs (B) TYPE: nucleotide (C) NUMBER OF CHAINS: single (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: Genomic DNA 10 (ix) CHARACTERISTICS: (A) NAME / KEY: exon (B) LOCATION: 1..20 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8: ATAACTGTTG GATTTCTCAA 20 15 (2) INFORMATION FOR SEQ ID NO: 9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 660 base pairs (B) TYPE: nucleotide 20 (C) NUMBER OF CHAINS: single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: genomic DNA (ix) CHARACTERISTICS: (A) NAME / KEY: exon 25 (B) LOCATION: 1.660 ^^^^^^^^^^^^^^^^^^^ fc ^^^^^^^^^^^^^^^^^^^^^^ g ^^^^^^ ^^^ J ^ JJ ^ ^^^^^^^^^^^^ M ^ ^^ j ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ SEQ ID NO: 9: TCGTGTGTGA ACATCACAGG GTTTGTGGAT GCACTTAGAT GTTTGCAATG AGCACTSTGG 60 CTGGCATGCC CCAGTGTTTT GGATACCAAT GCATAGGACT CCATAGTAAT CGAATTTACC 120 AGAGGCGAAC GTCATGAGCA TAGTGATCCC ATTGGGGGTT GATACAGCAG AGACGTCATA 180 CTTGGAAATG GCTGCAGGTT CAGAACCAGA ATCCGTAGAA GCTAGCCCTG TGGTAGTTGA 240 GAAATCCAAC AGTTATCCCC ACCAGTTATA TACCAGCAGC TCACATCATT CACACAGTTA 300 CATTGGTTTG CCCTATGCGG ACCATAATTA TGGTGCTCGT CCTC TCCGA CAC T CGGC 36C TTCCCCTCCT CCATCAGTCC TTATTAGCAA AAATGAAGTA GGCATATTTA CCACTCCTAA 420 TTTTGATGAA ACTTCCAGTG CTACTACAAT CA3CACATCT GAGGATGGAA GT ATGGTAC 480 TGATGTAACC AGGTGCATAT GTGGTTTTAC ACATGATGAT GGATACATGA TC 3TTGTGA 540 CAAATGCAGC GTTTGGCAAC ATATTGACTG CATGGGGATT GATAGGCAGC ATATTCCTGA 600 TACATATCTA TGTGAACGTT GTCAGCCTAG GAATTTGGAT AAAGAGAGGG CAGTGCTACT 660 (2) INFORMATION FOR SEQ ID NO: 10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 175 base pairs (B) TYPE: amino acid (C) NUMBER OF CHAINS: single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (ix) CHARACTERISTICS: (A) NAME / KEY: peptide (B) LOCATION: 1..175 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10: Met Ser lie Val lie Pro Leu Gly Val Asp Thr Ala Glu Thr Ser Tyr 5 10 15 Leu Glu Met Wing Wing Gly Ser Glu Pro Glu Ser Val Glu AJ.a Ser Pro 20 25 30 Val Val Val Glu Lys Ser Asn Ser Tyr Pro His Gln Leu Tyr Thr Ser 35 40 45 Uug ^ gUu &j ^ jjig Ser Ser His H¿s Ser His Ser Tyr He Gly Leu Pro Tyr Wing Asp Hxs 50 55 60 Asn Tyr Giy Wing Arg Pro Pro Pro Thr Pro Pro Wing Pro Pro Pro 65 7C "75 80 Ser Val Leu He Ser Lys Asn Glu Val Gly He Phe Thr Pro Asn 85 90 95 Phe Asp Glu Thr Ser Be Wing Thr Thr Ser Ser Thr Ser Glu Asp Gly 100 105 110 Ser Tyr Gly Thr Asp Val Thr Arg Cys He Cys Gly Phe Thr Hi = A = p 115 120 125 Asp Gly Tyr Met He Cys Cys Asp Lys Cys Ser Val Trp Gln HIS He 130 135 140 Asp Cys Met Gly He Asp Arg Gln His He Pro Asp Thr Tyr Leu Cys 145 150 155 160 Glu Arg Cys Gln Pro Arg Asn Leu Asp Lys Glu Arg Wing Val Leu 165 170 175 fifteen twenty lftatíüít.i ^^^ & giÉ ^ 7 j

Claims (1)

1. CLAIMS 1. An angiotensin receptor having at least one subunit containing the amino acid sequence SEQ ID NO: 10. The angiotensin receptor according to claim 1, wherein the amino acid sequence SEQ ID NO. 10 may possibly possess additional amino acids at the C-terminus. 3. The angiotensin receptor according to one or more of claims 1 and 2, wherein the subunit containing the sequence SEQ ID NO. 10 has a transmembrane domain. 4. The angiotensin receptor according to one or more of claims 1 to 3, wherein the receptor 15 binds to angiotensin 1-7 or to an angiotensin derivative 1-7. The angiotensin receptor according to one or more of claims 1 to 4, wherein at least one subunit of the receptor is encoded by an acid 20 nucleic having the sequence SEQ ID NO. 9, and where the sequence SEQ ID NO. 9 possibly contains additional nucleotides belonging to the sequence of; coding, at the 3 'end. 6. A process for the preparation of a nucleic acid that; 25 has the sequence SEQ ID NO. 9, where, in a reaction in a polymerase chain, the oligonucleotides whose sequences correspond to homologous regions in the nucleotide sequences of the angiotensin ATI and AT2 receptors are used as primers and where cDNA prepared from cells in which no receptors are expressed is used as a template ATI and AT2. A process for the preparation of an angiotensin receptor according to one or more of claims 1 to 5, wherein a nucleic acid which; contains the sequence SEQ ID NO. 9 is integrated into a suitable vector, the recombinant vector is introduced into a cell and the angiotensin receptor is expressed in this cell. The use of a nucleic acid containing the sequence SEQ ID NO. 9 in a process for the preparation of a recombinant angiotensin receptor, with the nucleic acid, which is under the control of a suitable promoter, introduced into a cell and expressed in this cell. The use of a nucleic acid containing the sequence SEQ ID NO. 9 to prepare a recombinant cell in which the angiotesin receptor can be expressed. The use of a nucleic acid having the sequence SEQ ID NO. 9 to produce a drug for treatment ^^ a & lfe. fe | g || Ég gg! ^^ and / or the prevention of diseases accompanied by defective regulation of the cardiovascular system, electrolyte balance and / or fluid balance. 11. The use of a nucleic acid containing the sequence SEQ ID NO. 9 to produce a drug that can be used in gene therapy. 12. The use of a nucleic acid containing the sequence SEQ ID NO. 9 in a process to identify and characterize substances that can be used as angiotensin receptor agonists or antagonists 13. The use of an angiotensin receptor according to one or more of claims 1 to 5 in a process for identifying and characterizing angiotensin receptor agonists or antagonists. 14. A process for identifying and characterizing angiotensin receptor agonists or antagonists where a nucleic acid having the sequence SEQ ID NO. 9 is introduced into a cell and the angiotensin receptor is expressed in this cell, this cell is incubated with a substance to be investigated and the effect of this substance on the angiotensin receptor is determined. 15. A drug comprising a nucleic acid having the sequence SEQ ID NO. 9. 16. A recombinant cell that is prepared by means of a nucleic acid having the sequence SEQ ID NO. 9 introduced in this cell and expressed in the cell. ¿^^^^^^^^^^^^^^^^^^^^, ^^^^^^^