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虹鳟(Oncorhynchus mykiss)肉碱棕榈酰转移酶I基因的克隆及组织分布

Cloning and tissue distribution of a carnitine palmitoyltransferase I gene in rainbow trout (Oncorhynchus mykiss).

作者信息

Gutières Stéphanie, Damon Marie, Panserat Stéphane, Kaushik Sadasivam, Médale Françoise

机构信息

Fish Nutrition Laboratory, Unité mixte INRA-IFREMER, Saint-Pée-sur-Nivelle 64310, France.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2003 May;135(1):139-51. doi: 10.1016/s1096-4959(03)00074-5.

Abstract

The carnitine palmitoyltransferase I (EC.2.3.1.21; CPT I) mediates the transport of fatty acids across the outer mitochondrial membrane. In mammals, there are two different proteins CPT I in the skeletal muscle (M) and liver (L) encoded by two genes. The carnitine palmitoyltransferase system of lower vertebrates received little attention. With the aim of improving knowledge on the CPT family in fish, we examined CPT I cDNA and CPT activity in different tissues of rainbow trout (Oncorhynchus mykiss). Using RT-PCR, we successfully cloned a partial CPT I cDNA sequence (1650 bp). The predicted protein sequence revealed identities of 63% and 61% with human L-CPT I and M-CPT I, respectively. This mRNA is expressed in liver, white and red skeletal muscles, heart, intestine, kidney and adipose tissue of trout. This is in good agreement with the measurement of the CPT activity in the same tissues. The [IC(50)] that reflects the sensitivity to malonyl-CoA inhibition was 0.116+/-0.004 microM for the liver and 0.426+/-0.041 microM for the white muscle. These results demonstrate for the first time the existence of at least one gene encoding for CPT I present in both the liver and the muscle of rainbow trout.

摘要

肉碱棕榈酰转移酶I(EC.2.3.1.21;CPT I)介导脂肪酸穿过线粒体外膜的转运。在哺乳动物中,由两个基因编码的骨骼肌(M)和肝脏(L)中有两种不同的CPT I蛋白。低等脊椎动物的肉碱棕榈酰转移酶系统很少受到关注。为了增进对鱼类CPT家族的了解,我们检测了虹鳟(Oncorhynchus mykiss)不同组织中的CPT I cDNA和CPT活性。利用逆转录聚合酶链反应(RT-PCR),我们成功克隆了一段CPT I cDNA序列(1650 bp)。预测的蛋白质序列与人类肝脏型CPT I和肌肉型CPT I的同源性分别为63%和61%。该mRNA在虹鳟的肝脏、白色和红色骨骼肌、心脏、肠道、肾脏及脂肪组织中表达。这与相同组织中CPT活性的检测结果高度一致。反映对丙二酰辅酶A抑制敏感性的半数抑制浓度(IC50),肝脏为0.116±0.004微摩尔,白色肌肉为0.426±0.041微摩尔。这些结果首次证明,虹鳟的肝脏和肌肉中至少存在一个编码CPT I的基因。

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