Paul R V, Wackym P S, Budisavljevic M, Everett E, Norris J S
Division of Nephrology, Ralph H. Johnson Department of Veterans Affairs Medical Center, Charleston, South Carolina.
J Biol Chem. 1993 Aug 25;268(24):18205-12.
Rat mesangial cells can express both 130-kDa guanylyl cyclase-coupled and 66-kDa non-coupled atrial natriuretic peptide (ANP) receptors (ANPR-A and ANPR-C, respectively). Exposure of mesangial cells, grown in 20% fetal calf serum, to 0.1% serum for 24 h increased total ANP receptor density more than 2-fold (Bmax = 87 versus 37 fmol/mg of cell protein) without changing binding affinity (Kd = 94 versus 88 pM). Radioligand binding and cross-linking studies demonstrated that up-regulation of ANP binding after serum deprivation was entirely due to an increase in ANPR-C, with little or no change in ANPR-A. Inhibition of protein synthesis with cycloheximide blocked up-regulation after serum deprivation. Steady-state ANPR-C mRNA level was increased 15-fold by serum deprivation, as judged by Northern blotting. There was no change in ANPR-A mRNA. Platelet-derived growth factor and phorbol myristate acetate, when added to low serum medium, blocked or reversed the effect of serum deprivation on ANPR-C. We conclude that synthesis and expression of ANPR-C but not ANPR-A is suppressed by serum, platelet-derived growth factor, and phorbol myristate acetate. Suppression of ANPR-C in vivo could contribute to mesangial cell proliferative responses to growth factors.
大鼠系膜细胞可表达130 kDa鸟苷酸环化酶偶联型和66 kDa非偶联型心房利钠肽(ANP)受体(分别为ANPR - A和ANPR - C)。将生长于20%胎牛血清中的系膜细胞置于0.1%血清中培养24小时,可使ANP受体总密度增加2倍以上(Bmax = 87对37 fmol/mg细胞蛋白),而结合亲和力不变(Kd = 94对88 pM)。放射性配体结合和交联研究表明,血清剥夺后ANP结合的上调完全归因于ANPR - C的增加,而ANPR - A几乎没有变化。用环己酰亚胺抑制蛋白质合成可阻断血清剥夺后的上调。通过Northern印迹法判断,血清剥夺使稳态ANPR - C mRNA水平增加了15倍。ANPR - A mRNA没有变化。当将血小板衍生生长因子和佛波醇肉豆蔻酸酯添加到低血清培养基中时,可阻断或逆转血清剥夺对ANPR - C的影响。我们得出结论,血清、血小板衍生生长因子和佛波醇肉豆蔻酸酯可抑制ANPR - C而非ANPR - A的合成和表达。体内ANPR - C的抑制可能有助于系膜细胞对生长因子的增殖反应。