Raynor K, Coy D C, Reisine T
Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia 19104.
J Neurochem. 1992 Oct;59(4):1241-50. doi: 10.1111/j.1471-4159.1992.tb08433.x.
Somatostatin (SRIF) is a neurotransmitter that produces its multiple effects in the CNS through interactions with membrane-bound receptors. Subtypes of SRIF receptors are found in the CNS that are distinguished by their sensitivities to the cyclic hexapeptide MK-678, such that SRIF1 receptors are sensitive to MK-678 and SRIF2 receptors are insensitive to MK-678. In the present study, we further examined the selectivities of a series of structurally diverse SRIF analogues for SRIF receptor subtypes. SRIF receptors were labeled by 125I-Tyr11-SRIF, which has indistinguishable affinities for SRIF receptor subtypes. The inhibition by MK-678 was incomplete, indicating this peptide is highly selective for a subtype of SRIF receptor that we have termed the SRIF1 receptor. The binding of 125I-MK-678 to SRIF1 receptors was monophasically inhibited by SRIF, the octapeptides (such as SMS-201-995), and the hexapeptides (such as MK-678), consistent with the highly selective labeling of a subtype of SRIF receptor. In contrast, the smaller CGP-23996-like analogues did not inhibit 125I-MK-678 binding to SRIF1 receptors. The binding of 125I-CGP-23996 to SRIF receptors was inhibited by SRIF and the octapeptides with Hill coefficients of less than 1, indicating that 125I-CGP-23996 labels multiple SRIF receptor subtypes. The hexapeptides and CGP-23996-like compounds produced only partial inhibitions of 125I-CGP-23996 binding, which were additive, indicating selective interactions of these compounds with the different receptor subpopulations labeled by 125I-CGP-23996. 125I-Tyr11-SRIF binding and 125I-CGP-23996 binding to SRIF receptors were likewise only partially affected by 100 microM guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), a concentration that completely abolishes specific 125I-MK-678 binding to SRIF1 receptors. The component of 125I-CGP-23996 labeling that was sensitive to GTP gamma S was also MK-678 sensitive. Thus, two subpopulations of SRIF receptors exist in the CNS. The SRIF1 receptor is sensitive to cyclic hexapeptides such as MK-678 and to GTP gamma S but insensitive to smaller CGP-23996-like compounds. The SRIF2 receptor is sensitive to the CGP-23996-like compounds and can be selectively labeled by 125I-CGP-23996 in the presence of high concentrations of the hexapeptides or GTP gamma S because, unlike the SRIF1 receptor, the SRIF2 receptor is insensitive to these agents. The SRIF receptor subtype-selective peptide analogues will be useful in the future characterization of the functions mediated by SRIF receptor subtypes in the CNS.
生长抑素(SRIF)是一种神经递质,它通过与膜结合受体相互作用在中枢神经系统中产生多种效应。在中枢神经系统中发现了SRIF受体亚型,它们对环状六肽MK - 678的敏感性不同,即SRIF1受体对MK - 678敏感,而SRIF2受体对MK - 678不敏感。在本研究中,我们进一步研究了一系列结构多样的SRIF类似物对SRIF受体亚型的选择性。SRIF受体用125I - Tyr11 - SRIF标记,它对SRIF受体亚型具有难以区分的亲和力。MK - 678的抑制作用不完全,表明该肽对我们称为SRIF1受体的SRIF受体亚型具有高度选择性。125I - MK - 678与SRIF1受体的结合被SRIF、八肽(如SMS - 201 - 995)和六肽(如MK - 678)单相抑制,这与SRIF受体亚型的高度选择性标记一致。相比之下,较小的CGP - 23996样类似物不抑制125I - MK - 678与SRIF1受体的结合。125I - CGP - 23996与SRIF受体的结合被SRIF和八肽抑制,希尔系数小于1,表明125I - CGP - 23996标记多种SRIF受体亚型。六肽和CGP - 23996样化合物仅对125I - CGP - 23996结合产生部分抑制,且这些抑制作用是相加的,表明这些化合物与125I - CGP - 23996标记的不同受体亚群有选择性相互作用。125I - Tyr11 - SRIF结合和125I - CGP - 23996与SRIF受体的结合同样仅受到100 microM鸟苷5'-O -(3 - 硫代三磷酸)(GTPγS)的部分影响,该浓度可完全消除125I - MK - 678与SRIF1受体的特异性结合。对GTPγS敏感的1部分125I - CGP - 23996标记也对MK - 678敏感。因此,中枢神经系统中存在两种SRIF受体亚群。SRIF1受体对环状六肽如MK - 678和GTPγS敏感,但对较小的CGP - 23996样化合物不敏感。SRIF2受体对CGP - 23996样化合物敏感,并且在高浓度六肽或GTPγS存在下可被125I - CGP - 23996选择性标记,因为与SRIF1受体不同,SRIF2受体对这些试剂不敏感。SRIF受体亚型选择性肽类似物将有助于未来对中枢神经系统中SRIF受体亚型介导的功能进行表征。