ANGIOTENSIN-IV RECEPTORS AND SIGNALING IN OPOSSUM KIDNEY-CELLS

被引:51
作者
DULIN, N
MADHUN, ZT
CHANG, CH
BERTIMATTERA, L
DICKENS, D
DOUGLAS, JG
机构
来源
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL FLUID AND ELECTROLYTE PHYSIOLOGY | 1995年 / 269卷 / 05期
关键词
OK7A CELL; GUANOSINE 5'-O-(3-THIOTRIPHOSPHATE); DITHIOTHREITOL; GUANOSINE; 3'; 5'-CYCLIC MONOPHOSPHATE; ADENOSINE; PHOSPHOLIPASE C; CALCIUM;
D O I
10.1152/ajprenal.1995.269.5.F644
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
The pharmacological properties and signaling of angiotensin TV (ANG TV) receptors were studied in opossum kidney cell line OK7A. Saturation binding experiments with I-125-labeled ANG IV demonstrated the presence of high-affinity ANG IV binding sites in OK7A cell membranes with a dissociation constant (K-d) Of 0.40 +/- 0.08 nM and a maximal amount of binding sites (B-max) of 180 +/- 50 fmol/mg protein. In competition experiments, unlabeled ANG TV inhibited I-125-ANG IV binding biphasically: 20% of binding sites had high affinity [inhibition constant (K-i) = 0.44 +/- 0.04 nhl] and 80% had low affinity (K-i = 130 +/- 10 nM). ANG III displaced I-125-ANG IV from binding sites with low affinity (K-i = 205 +/- 10 nM), and ANG II did not compete with I-125-ANG IV at concentrations up to 10 mu M. The binding of ANG IV to OK7A cell membranes was significantly enhanced in the presence of 5 mM EDTA and completely blocked by 5 mM dithiothreitol. Guanosine 5'-O-(3-thiotriphosphate) inhibited the binding of I-125-ANG IV, indicating the G protein coupling of ANG IV receptors in OK7A cells. In signaling studies, ANG IV induced transient increase in intracellular calcium concentration ([Ca2+](i)) from 49 +/- 3 to 280 +/- 45 nM. ANG IV failed to influence phosphoinositol metabolism, indicating that Ca2+ mobilization is not linked to ANG IV signaling. Ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid completely abolished ANG IV-induced increase in [Ca2+](i), consistent with Ca2+ influx. The voltage-sensitive Ca2+ channel blocking agents verapamil and nifedipine attenuated the effect of ANG IV on [Ca2+](i) to 133 +/- 33 and 174 +/- 32 nM, respectively. These data suggest that ANG TV induces Ca2+ influx in OK7A cells, at least partially, through the voltage-sensitive Ca2+ channels.
引用
收藏
页码:F644 / F652
页数:9
相关论文
共 44 条
  • [1] ANGIOTENSIN-II STIMULATION OF PROTEIN-SYNTHESIS AND CELL-GROWTH IN CHICK HEART-CELLS
    BAKER, KM
    ACETO, JF
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1990, 259 (02): : H610 - H618
  • [2] A SPECIFIC BINDING-SITE RECOGNIZING A FRAGMENT OF ANGIOTENSIN-II IN BOVINE ADRENAL-CORTEX MEMBRANES
    BERNIER, SG
    FOURNIER, A
    GUILLEMETTE, G
    [J]. EUROPEAN JOURNAL OF PHARMACOLOGY, 1994, 271 (01) : 55 - 63
  • [3] EFFECT OF HEPTAPEPTIDE (2-8) AND HEXAPEPTIDE (3-8) FRAGMENTS OF ANGIOTENSIN-II ON ALDOSTERONE SECRETION
    BLAIRWES.JR
    COGHLAN, JP
    DENTON, DA
    FUNDER, JW
    SCOGGINS, BA
    WRIGHT, RD
    [J]. JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM, 1971, 32 (04) : 575 - +
  • [4] ANGIOTENSIN-II-(3-8)-HEXAPEPTIDE AFFECTS MOTOR-ACTIVITY, PERFORMANCE OF PASSIVE-AVOIDANCE AND A CONDITIONED AVOIDANCE-RESPONSE IN RATS
    BRASZKO, JJ
    KUPRYSZEWSKI, G
    WITCZUK, B
    WISNIEWSKI, K
    [J]. NEUROSCIENCE, 1988, 27 (03) : 777 - 783
  • [5] ANGIOTENSIN-II STIMULATION OF NA-H ANTIPORTER ACTIVITY IS CAMP-INDEPENDENT IN OKP CELLS
    CANO, A
    MILLER, RT
    ALPERN, RJ
    PREISIG, PA
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1994, 266 (06): : C1603 - C1608
  • [6] CHANG CH, 1993, J BIOL CHEM, V268, P4908
  • [7] DISCRIMINATION OF ANGIOTENSIN-II RECEPTOR SUBTYPES BY DITHIOTHREITOL
    CHIU, AT
    MCCALL, DE
    NGUYEN, TT
    CARINI, DJ
    DUNCIA, JV
    HERBLIN, WF
    UYEDA, RT
    WONG, PC
    WEXLER, RR
    JOHNSON, AL
    TIMMERMANS, PBMWM
    [J]. EUROPEAN JOURNAL OF PHARMACOLOGY, 1989, 170 (1-2) : 117 - 118
  • [8] REGULATION OF CYTOSOLIC CALCIUM BY ANGIOTENSINS IN VASCULAR SMOOTH-MUSCLE
    DOSTAL, DE
    MURAHASHI, T
    PEACH, MJ
    [J]. HYPERTENSION, 1990, 15 (06) : 815 - 822
  • [9] DOUGLAS JG, 1994, ANNU REV PHYSIOL, V56, P649
  • [10] RABBIT RENAL EPITHELIAL ANGIOTENSIN-II RECEPTORS
    DULIN, NO
    ERNSBERGER, P
    SUCIU, DJ
    DOUGLAS, JG
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-RENAL FLUID AND ELECTROLYTE PHYSIOLOGY, 1994, 267 (05): : F776 - F782