Identification of LDL-receptor transcription stimulating peptides from soybean hydrolysate in human hepatocytes
Identification of LDL-receptor transcription stimulating peptides from soybean hydrolysate in human hepatocytes
复制标题
DOI：
10.1021/jf800676a
复制
发表时间：
2008-06-25
影响因子：
6.1
通讯作者：
Lee, Cherl-Ho
中科院分区：
文献类型：
--
作者：
Cho, Seong-Jun;Juillerat, Marcel A.;Lee, Cherl-Ho
据报道，大豆蛋白及其水解产物具有降低胆固醇的特性，但其具体成分仍不清楚。在之前的研究中，我们发现用解淀粉芽孢杆菌FSE-68的蛋白酶制备的大豆蛋白水解物（SPH）能够强烈刺激低密度脂蛋白受体（LDL-R）的转录。为了鉴定人肝细胞中的 LDL-R 转录刺激肽，使用凝胶渗透色谱法对 SPH 进行分级，并使用反相色谱法进一步分离活性部分。通过 LC/MS 和 MS/MS 分析鉴定了最活跃级分中的几种肽。根据已鉴定的序列合成了LDL-R转录刺激肽，并在体外测试了它们对LDL-R转录的影响。在合成的肽中，Phe-Val-Val-Asn-Ala-Thr-Ser-Asn（FVVNATSN）表现出最强的活性，并且FVVNATSN在100μM浓度下将肝细胞的LDL-R转录增加至248.8％（与未处理对照的100％相比）。本研究提供了大豆蛋白衍生肽可以影响LDL-R的直接证据。 肝细胞中的转录。
Soybean protein and its hydrolysate have been reported to have cholesterol-lowering property, but the responsible components are still largely unknown. In previous study, we found that soybean protein hydrolysate (SPH) prepared with the protease from Bacillus amyloliquefaciens FSE-68, strongly stimulates transcription of low density lipoprotein receptor (LDL-R). To identify LDL-R transcription stimulating peptides in human hepatocytes, the SPH was fractionated with gel permeation chromatograpy and the active fraction was further separated by using reverse-phase chromatography. Several peptides in the most active fraction were identified by LC/MS and MS/MS analysis. LDL-R transcription stimulating peptides were synthesized on the basis of identified sequences, and their effect on LDL-R transcription was tested in vitro. Among the synthesized peptides, Phe-Val-Val-Asn-Ala-Thr-Ser-Asn (FVVNATSN) showed the strongest activity, and LDL-R transcription of hepatic cells was increased to 248.8% (compared to 100% of untreated control) by FVVNATSN at a concentration of 100 mu M. This study provides direct evidence that peptides derived from soybean protein can influence LDL-R transcription in hepatocytes.