Molecular cloning, characterization, and distribution of the gerbil angiotensin II AT2 receptor.
We isolated a cDNA clone encoding the gerbil AT2 receptor (gAT2) gene from a gerbil adrenal gland cDNA library. The full-length cDNA contains a 1,089-bp open reading frame encoding 363 amino acid residues with 90.9, 96.1, and 95.6% identity with the human (hAT2), rat (rAT2), and mouse AT2 (mAT2) receptors, respectively. There are at least seven nonconserved amino acids in the NH2-terminal domain and in positions Val196, Val217, and Met293, important for angiotensin (ANG) II but not for CGP-42112 binding. Displacement studies in adrenal sections revealed that affinity of the gAT2 receptor was 10-20 times lower for ANG II, ANG III, and PD-123319 than was affinity of the rAT2 receptor. The affinity of each receptor remained the same for CGP-42112. When transfected into COS-7 cells, the gAT2 receptor shows affinity for ANG II that is three times lower than that shown by the hAT2 receptor, whereas affinities for ANG III and the AT2 receptor ligands CGP-42112 and PD-123319 were similar. Autoradiography in sections of the gerbil head showed higher binding in muscles, retina, skin, and molars at embryonic day 19 than at 1 wk of age. In situ hybridization and emulsion autoradiography revealed that at embryonic day 19 the gAT2 receptor mRNA was highly localized to the base of the dental papilla of maxillary and mandibular molars. Our results suggest selective growth-related functions in late gestation and early postnatal periods for the gAT2 receptor and provide an essential basis for future mutagenesis studies to further define structural requirements for agonist binding.
Hoe
Kwang-Lae
KL
Section on Pharmacology, National Institute of Mental Health, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland 20892-1514, USA. armandoi@intra.nimh.nih.gov
Armando
Ines
I
Baiardi
Gustavo
G
Sreenath
Taduru
T
Kulkarni
Ashok
A
Martínez
Alfredo
A
Saavedra
Juan M
JM
D000311:Adrenal Glands / Q000196:embryology / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D019556:COS Cells; D003001:Cloning, Molecular; D018507:Gene Expression Regulation, Developmental; D005849:Gerbillinae; D006257:Head / Q000196:embryology / Q000502:physiology*; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000032:analysis; D011869:Radioligand Assay; D051381:Rats; D044139:Receptor, Angiotensin, Type 2 / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity; D014070:Tooth / Q000196:embryology / Q000502:physiology*; D014162:Transfection
D012333:RNA, Messenger; D044139:Receptor, Angiotensin, Type 2