小肽转运载体(PepT1和PepT2)研究进展

由小肽转运载体介导的小肽吸收对动物的生长和发育有着特殊的作用,因此,对小肽转运载体的深入研究在生理、药理和临床上都有着非常重要的意义。文中主要从小肽转运载体(PepT1和PepT2)的蛋白质分子结构与功能、主要的组织分布和影响其活性的因素三方面进行简要综述。     

小肽转运载体2的转运机制及功能研究

小肽转运载体2(peptide transporter 2,PepT2)是一种高亲和力、低容量的转运蛋白,能转运大多数小肽类营养物质和仿肽类药物,因此,对PepT2进行深入研究对动物营养学和医学临床治疗均具有重要意义。本文综述了PepT2的功能结构、转运机制及其底物结合特性,阐述了其在不同组织中的功能及活性调节,并对其今后的研究方向进行了展望。     

小肽转运载体2在奶牛乳腺小肽摄取中的作用研究

本试验旨在研究2型小肽转运载体(oligopeptide transporter 2,PepT 2)在奶牛乳腺组织吸收利用小肽合成乳蛋白过程中的作用.在体外培养的奶牛乳腺组织培养液中分别添加不同浓度的苯丙氨酸二肽( Phe-Phe)(0和11.7 μg/mL)和/或焦碳酸二乙酯(DEPC)(0、0.01、0.1、0.5...     

科技前沿

小肽转运载体2在奶牛乳腺小肽摄取中的作用研究试验旨在研究2型小肽转运载体(oligopeptide transporter 2,PepT2)在奶牛乳腺组织吸收利用小肽合成乳蛋白过程中的作用。在体外培养的奶牛乳腺组织培养液中分别添加不同浓度的苯丙氨酸二肽(Phe-Phe)(0和11.7μg/mL)和/或焦碳酸二乙酯(DEPC)(0、0.01、0.1、0.5和1.0mmol/L)进行培养,试验结束后     

小肽转运载体1的生物学特性及其功能

小肽转运载体1(PepT1)是H+/肽偶联的转运载体。该载体通过利用肠腔到肠细胞的质子梯度来转运二肽和三肽。PepT1对游离氨基酸、多肽在动物肠道内的转运调控具有重要作用。本文综述了PepT1的分类、生物学特征及功能,并探讨了影响PepT1活性调控的因素。     

小肽转运蛋白(PepT1)基因研究进展

小肽作为蛋白质的主要消化产物 ,在氨基酸消化、吸收和代谢中起着重要作用。小肽与游离氨基酸的吸收是两个相互独立的转运系统 ,与游离氨基酸相比 ,小肽具有吸收速度快、耗能低、不易饱和 ,且各种肽之间转运无竞争性与抑制性等特点。本文主要综述了小肽转运机制的特点和小肽转运蛋白(PepT1)分子生物学方面的研究进展 ,包括PepT1分子结构特点 ,PepT1cDNA的克隆 ,不同动物之间PepT1氨基酸序列的同源性 ,PepT1mRNA在不同动物、不同组织中的分布 ,以及营养水平对PepT1基因表达的影响 ;并就需要进一步深入研究的问题进行了探讨     

寡肽转运蛋白(PepT1)生物学特性及活性调控研究进展

作为蛋白质主要的消化产物,小肽的吸收不仅在氨基酸消化、吸收和代谢中起到至关重要的作用,对于维持反刍动物的生长、发育也具有重要意义。目前寡肽转运蛋白(PepT1)被预测出有12个跨膜结构域,Fei等对其跨膜结构域进行了初步的研究。PepT1所具有的无从差别转运小肽、在特定的PH条件下电现象等特点也使得其成为学者们研究的热点。此外,底物因素、生理生物因素以及营养因素等对于PepT1活性调控的影响也是人们研究的热点。本文基于对PepT1生物学特性及活性调控研究进展的论述,对PepT1的研究前景进行了展望,以期为促进PepT1的进一步研究提供进一步的参考。     

反刍动物瘤胃物质转运系统及其调控的研究

小肽、挥发性脂肪酸（VFA）和葡萄糖是反刍动物体内重要的营养物质．对反刍动物的生产性能和产品质量影响重大。研究发现小肽的主要转运载体有两个．PepT1和PepT2：葡萄糖的转运载体分两大类共16个载体；VFA的瘤胃转运主要受PH、日粮精粗比、分子大小及血液流量的调控。本文对此3种重要营养物质在反刍动物瘤胃内的转运机制和调控因子进行综述．并展望以后的研究动向.     

鱼类小肽转运载体PepT1研究进展

小肽作为蛋白质主要的消化产物,与氨基酸的吸收是两个相互独立的转运系统。Peptide Transporter 1(PepT1)是其中一种重要的小肽转运载体。文章就鱼类PepT1的分子特点,PepT1 cDNA的克隆与表达,以及PepT1的转运机制等研究作一综述。     

小肽转运蛋白(PepT1)的活性调节

小肽是蛋白质在动物体内消化的主要产物,而位于小肠刷状缘膜上的小肽转运蛋白(PepT1)在小肽的吸收过程中发挥着重要作用,因此PepT1的活性直接影响小肽的吸收。文中主要介绍底物、营养不良、激素、昼夜节律和生长发育对PepT1的活性调节。     

Cloning,tissue distribution and functional characterization of the donkey (Equus asinus) oligopeptide transporter 2

Oligopeptide transporter 2 (PepT2) is an important transporter of oligopeptides. In the present study, we describe the molecular cloning, tissue distribution and functional characterization of a donkey (Equus asinus) PepT2. The cloned cDNA sequence was 2202 bp at full length, encoding a 733 amino acid peptide with a molecular weight of 81.9 kDa and a theoretical pI of 8.92. Bioinformatics analysis showed that the deduced peptide sequence possessed all the characteristic features of PepT2. The expression of PepT2 in the kidney and lung was significantly higher than that observed in the ileum, duodenum, jejunum, spleen, liver, heart and stomach. Functional characterization by heterologous expression in Chinese hamster ovary cells showed that the uptake of β-Ala-Lys-N-7-amino-4-methylcoumarin-3-acetic acid (β-Ala-Lys-AMCA) by donkey PepT2-Chinese hamster ovary cells was dependent on time, pH and substrate concentration, with a low Km value of 91.51 ± 14.14 μM and a maximum velocity of 41.37 ± 2.193 pmol/min/mg protein. In the present study, for the first time, the expression and functional characteristics of donkey PepT2 were evaluated, the results of which provide new insights and a better understanding of its crucial role in oligopeptide transport in donkeys.     

Effects of tripeptides and lactogenic hormones on oligopeptide transporter 2 in bovine mammary gland

This study was conducted to investigate the expression of oligopeptide transporter 2 (PepT2) and its potential function in bovine mammary gland. First, the PepT2 mRNA and protein were determined in cultured mammary epithelial cells. Then the effects of lactogenic hormones (prolactin, hydrocortisone or insulin) and substrate (threonyl-phenylalanyl-phenylalanine) on PepT2 were investigated. The PepT2 mRNA and protein were successfully detected in bovine mammary epithelial cells. PepT2 gene expression was enhanced by the addition of 50, 500 and 5000 ng/ml prolactin, 10 and 100 ng/ml hydrocortisone, and 50, 500, 5000 and 50,000 ng/ml insulin. PepT2 mRNA abundance was increased when 5, 10 and 15% of threonyl-phenylalanyl-phenylalanine was included. Responses of PepT2 to lactogenic hormones and oligopeptide inferred that it may play an important role in bovine mammary gland.     

Board-invited review: Peptide absorption and utilization: Implications for animal nutrition and health

Over the last 50 yr, the study of intestinal peptide transport has rapidly evolved into a field with exciting nutritional and biomedical applications. In this review, we describe from a historical and current perspective intestinal peptide transport, the importance of peptides to whole-body nutrition, and the cloning and characterization of the intestinal peptide transporter, PepT1. We focus on the nutritional significance of peptide transport and relate these findings to livestock and poultry. Amino acids are transported into the enterocyte as free AA by a variety of AA transporters that vary in substrate specificity or as di- and tripeptides by the peptide transporter, PepT1. Expression of PepT1 is largely restricted to the small intestine in most species; however, in ruminants, peptide transport and activity is observed in the rumen and omasum. The extent to which peptides are absorbed and utilized is still unclear. In ruminants, peptides make a contribution to the portal-drained visceral flux of total AA and are detected in circulating plasma. Peptides can be utilized by the mammary gland for milk protein synthesis and by a variety of other tissues. We discuss the factors known to regulate expression of PepT1 including development, diet, hormones, diurnal rhythm, and disease. Expression of PepT1 is detected during embryological stages in both birds and mammals and increases with age, a strategic event that allows for the immediate uptake of nutrients after hatch or birth. Both increasing levels of protein in the diet and dietary protein deficiencies are found to upregulate the peptide transporter. We also include in this review a discussion of the use of dietary peptides and potential alternate routes of nutrient delivery to the cell. Our goal is to impart to the reader the nutritional implications of peptide transport and dietary peptides and share discoveries that shed light on various biological processes, including rapid establishment of intestinal function in early neonates and maintenance of intestinal function during fasting, starvation, and disease states.     

Functional characterization of a cloned pig intestinal peptide transporter (pPepT1)

Absorption of dietary protein can be mediated through the uptake of AA as free AA or small peptides. A H(+)-coupled, peptide transport protein, PepT1, is responsible for the absorption of small peptides arising from digestion of dietary proteins in the small intestine. The magnitude of peptide absorption and the nutritional significance of PepT1 are unknown for many food-producing animals; thus, the objective of this study was to clone and determine the functional characteristics of the pig PepT1 (pPepT1). Two cDNA-encoding pPepT1 were isolated, which contain alternative polyadenylation sites. The predicted pPepT1 is a 708-AA protein, which shows 82.8, 85.7, and 64.7% AA identity to human, sheep, and chicken PepT1, respectively. On northern blots, two pPepT1 mRNA of approximately 2.9 and 3.5 kb were detected in the duodenum, jejunum, and ileum of the small intestine and are presumed to result from alternative polyadenylation. Uptake of [(3)H]-Gly-Sar was measured in Chinese hamster ovary cells transiently transfected with a pPepT1 expression vector to study the functional expression of pPepT1. Peptide transport was H(+)-dependent, with an optimal pH of 6.0 to 6.5. The ability of pPepT1 to transport various peptides was assayed by calculating the concentration of unlabeled peptide that inhibited 50% of [(3)H]-Gly-Sar uptake (IC(50)) in transfected cells. Eleven dipeptides and two tripeptides had IC(50) values that ranged from 0.004 to 0.53 mM. Three peptides, Lys-Lys, Arg-Lys, and Lys-Trp-Lys, had IC(50) values greater than 1. 38 mM and seem to be poor substrates for pPepT1. For all three tetrapeptides examined, uptake of Gly-Sar was too small to measure, even at a concentration of 10 mM tetrapeptide; therefore, IC(50) values could not be calculated. These results demonstrate that pPepT1 can transport a variety of dipeptides and tripeptides but not tetrapeptides.     

Expression of Proton-dependent Oligopeptide Transporters (POTs) in Calves' Tissues

This test was designed to study the expression of POTs (PepT1, PepT2, PHT1 and PHT2) mRNA in calves'tissues.Tissue-specific expression of the mRNA corresponding to peptide transporter protein in four 3-month-old Chinese Holstein calves was examined by relative quantitative RT-PCR analysis.The results showed that the expression of PepT1 mRNA in the rumen was extremely significantly higher than that in the heart and muscle (P<0.01);The expression of PepT2 mRNA in the liver and kidney was extremely significantly higher than that in the heart, spleen, thymus, muscle, rumen, reticulum, omasum and abomasum (P<0.01);The expression of PHT1 mRNA in the lung and spleen was extremely significantly higher than that in the heart and muscle (P<0.01);The expression of PHT2 mRNA in the lung and thymus was significantly higher than that in the heart, liver, kidney, spleen, muscle, rumen, reticulum, omasum and abomasum (P<0.05).The results indicated that PepT1, PepT2, PHT1 and PHT2 mRNA were respectively abundantly expressed in rumen, kidney and liver, spleen and lung, lung and thymus.     

犊牛组织中小肽转运体(POTs)的表达研究

为研究犊牛部分组织中小肽转运体(proton-dependent oligopeptide transporters,POTs) (PepT1、PepT2、PHT1、PHT2) mRNA的表达,试验选用4头3月龄的中国荷斯坦犊牛进行屠宰,采用实时荧光定量PCR方法来定量组织中小肽转运蛋白表达水平。结果表明,犊牛PepT1 mRNA在瘤胃中表达丰度极显著高于心脏和肌肉(P<0.01);犊牛PepT2 mRNA在肝脏和肾脏中表达丰度极显著高于心脏、脾脏、胸腺、肌肉、瘤胃、网胃、瓣胃、皱胃(P<0.01);犊牛PHT1 mRNA在肺脏和脾脏中表达丰度极显著高于心脏和肌肉(P<0.01);犊牛PHT2 mRNA在肺脏和胸腺中表达丰度显著高于心脏、肝脏、肾脏、脾脏、肌肉、瘤胃、网胃、瓣胃、皱胃(P<0.05)。综上所述,PepT1、PepT2、PHT1、PHT2 mRNA分别在犊牛瘤胃、肾脏和肝脏、脾脏和肺脏、肺脏和胸腺中表达量最高。     

肉鸡肠道PepT1 mRNA表达的肠段差异性与发育性变化

选用遗传背景相同的1日龄父母代雄性Arbor Acre(AA)鸡和父母代雄性岭南黄肉雏鸡各120羽,采用相对定量RT-PCR方法,以30 d时岭南黄鸡肠道样品为模板,研究肉鸡肠道寡肽转运载体1(Peptide transporter 1,PepT1)mRNA表达的肠段差异性;以AA肉鸡和岭南黄鸡十二指肠和空肠样品为模板,研究不同品种肉鸡肠道PepT1 mRNA表达的发育性变化。结果显示:(1)岭南黄肉鸡肠道PepT1 mRNA的表达丰度从十二指肠、空肠、回肠到结直肠依次降低,其中十二指肠显著高于结直肠(P<0.05);(2)AA鸡和岭南黄肉鸡PepT1 mRNA在十二指肠及空肠中的表达具有相同的发育模式,16 d表达丰度最高,16~44 d逐渐下降,58 d略微回升;不同基因型之间PepT1 mRNA丰度比较,AA鸡和岭南黄肉鸡两品种间PepT1 mRNA的表达没有显著差异(P>0.05)。以上结果说明:(1)随着肠道空间位置的后移,岭南黄肉鸡肠道PepT1 mRNA的表达丰度逐渐降低,其中十二指肠的表达丰度显著高于结直肠(P<0.05);(2)不同品种肉鸡十二指肠及空肠PepT1 mRNA的表达具有相同的发育模式,且同日龄两品种间的表达丰度未见明显差异,表明PepT1 mRNA表达受到发育阶段的调控,但品种间具有稳定性。