Billington A P, Walstrom K M, Ramesh D, Guzikowski A P, Carpenter B K, Hess G P
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
Biochemistry. 1992 Jun 23;31(24):5500-7. doi: 10.1021/bi00139a011.
Three photolabile precursors of glycine containing a photosensitive 2-nitrobenzyl moiety attached to the amino group have been synthesized. When exposed to ultraviolet radiation between 308 and 350 nm, the compounds photolyze to release glycine, an important inhibitory neurotransmitter in the central nervous system. The identification of glycine as a photolysis product was determined by two different methods: separation of the photolyzed sample by thin-layer chromatography followed by a reaction with ninhydrin, and recognition of derivatized glycine using the Waters pico-tag method in conjunction with high-performance liquid chromatography. The photolysis of these compounds at 22 degrees C has been investigated, and the rate of decay of a transient intermediate in the reaction, which is assumed to reflect product release, has been measured. For N-(alpha-carboxy-2-nitrobenzyl)glycine this decay rate was found to be 940 s-1 at pH 6.8 and 600 s-1 at pH 7.5. Additionally, this compound was found to exhibit biological activity upon photolysis; cultured mouse spinal cord cells containing neuronal glycine receptors were used to detect the glycine liberation. The approach adopted here is useful in demonstrating the utility of photolabile precursors of neurotransmitters that have the protecting group linked to the neurotransmitter through the amino group. The rapid photolysis of such compounds to release free neurotransmitter is valuable in gaining access to chemical kinetic studies of neurotransmitter receptors. Previously, such studies have been limited because the available methods for neurotransmitter delivery did not give a sufficiently high time resolution.
已合成了三种含甘氨酸的光不稳定前体,其氨基连接有光敏的2-硝基苄基部分。当暴露于308至350nm的紫外线辐射时,这些化合物发生光解以释放甘氨酸,甘氨酸是中枢神经系统中一种重要的抑制性神经递质。通过两种不同的方法确定甘氨酸为光解产物:通过薄层色谱法分离光解样品,然后与茚三酮反应,以及使用沃特世pico-tag方法结合高效液相色谱法识别衍生化的甘氨酸。已研究了这些化合物在22℃下的光解,并测量了反应中瞬态中间体的衰减速率,该衰减速率被认为反映了产物释放。对于N-(α-羧基-2-硝基苄基)甘氨酸,在pH 6.8时该衰减速率为940 s-1,在pH 7.5时为600 s-1。此外,发现该化合物在光解时表现出生物活性;使用含有神经元甘氨酸受体的培养小鼠脊髓细胞来检测甘氨酸的释放。这里采用的方法有助于证明神经递质光不稳定前体的实用性,这些前体的保护基团通过氨基与神经递质相连。此类化合物快速光解以释放游离神经递质,对于进行神经递质受体的化学动力学研究非常有价值。以前,此类研究受到限制,因为可用的神经递质递送方法没有给出足够高的时间分辨率。