ANALYSIS OF PROTEIN TYROSINE SULFATION

蛋白质酪氨酸硫酸化分析

• 批准号: 7723123
• 负责人: GRACE L ROSENQUIST
• 金额: $ 0.05万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7723123
• 关键词: 2-tyrosine; Achievement; Affect; Amino Acid Sequence; Amino Acids; Animals; Cells; Characteristics; Computer Retrieval of Information on Scientific Projects Database; Computer-Assisted Image Analysis; Consensus Sequence; Databases; Destinations; Endocrine; Evaluation; Funding; Glutamine; Goals; Golgi Apparatus; Grant; Inorganic Sulfates; Institution; Lead; Manuscripts; Numbers; Pattern; Peptide Sequence Determination; Positioning Attribute; Post-Translational Protein Processing; Process; Protein Analysis; Protein Sequence Analysis; Proteins; Publications; Range; Rattus; Research; Research Personnel; Resources; Score; Side; Site; Solvents; Source; SwissProt; System; Thinking; Tissues; Tyrosine; United States National Institutes of Health; Unspecified or Sulfate Ion Sulfates; aspartylglycine; base; glycylproline; in vivo; methionyltryptophan; protein function; protein-tyrosine sulfotransferase; research study; sulfation; sulfurtransferase; trafficking; tyrosine O-sulfate

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Sulfation is a widespread post-translational modification of tyrosines found in all animal cells containing a Golgi system. Our recent analysis of tyrosine sulfation sites showed that almost half contained the tyrosine trafficking motif, Y-x-x-Hydrophobic, (x being any amino acid). Since proteins containing either tyrosine-based site are processed in the trans-Golgi, what directs them to the correct destination? We have contrasted trafficking motifs with tyrosine sulfation sites, both containing the Y-x-x-hydrophobic pattern, to determine sequence features that distinguish these different tyrosine-based sites. Several log-odds position specific scoring matrices (PSSM) were created to distinguish sulfation sites from trafficking motifs. Sulfation sites and trafficking motifs were contrasted with each other, as well as against the Swiss-Prot metazoan database. Three of thirteen trafficking motifs scored in the range of known tyrosine sulfation sites. Evaluation of the PSSMs showed that the pattern of high-scoring amino acids differs for trafficking motifs and sulfation sites. In trafficking motifs, arg and gln are preferred at position y+1 and gly at y-1, whereas in sulfation sites small residues (asp, gly, pro) are preferred at y+1, and asp at y-1. Leu is preferred at y+3 in trafficking motifs, while met and trp are preferred in sulfation sites. These results indicate that although significant differences exist between tyrosine sulfation sites and trafficking motifs, there is some overlap between these two tyrosine-based sites. Further experiments should determine whether some trafficking motifs are sulfated in vivo, and whether sulfation affects a proteinsultimate destination. Additional factors such as additional motifs may be necessary to direct proteins to their correct destinations. In fact, approximately 1% of the tyrosines are sulfated in tissue in the rat. Despite this observation, only 24 unique protein sequences containing tyrosine sulfate have been described. The aim of this project is to analyze the amino acid sequences surrounding know tyrosine sulfation sites to determine the characteristics of this site and, therefore, to identify more candidates of sulfation. The specific goals are: 1. to identify the characteristics of sulfated tyrosines which distinguish them from non-sulfated tyrosines. These characteristics include the number and from non-sulfated tyrosines. These characteristics include the possible distribution of amino acids, as well as the possible secondary structural secondary structural features in the vicinity of the tyrosine. 2. to identify features in the vicinity of the tyrosine. 2. to identify proteins with probable proteins with probable tyrosine sulfation sites from the Swiss Protein data base. 3. to determine sequence similarities among the set of known proteins in tyrosine sulfation sites from the Swiss Protein data base. 3. to determine sequence similarities among the set of known proteins in the vicinity of the the vicinity of the sulfation site. Identification of new protein sulfation sites may be essential for the understanding of the function of these proteins sulfation site. Identification of new protein sulfation sites may be essential and will lead to a better understanding of the function of tyrosine sulfation. for the understanding of the function of these proteins and will lead to a better understanding of the function of tyrosine sulfation. In fact, approximately 1% of the tyrosines are sulfated in tissue in the rat. Despite this observation, only 24 unique protein sequences containing tyrosine sulfate have been described. The aim of this project is to analyze the amino acid sequences surrounding know tyrosine sulfation sites to determine the characteristics of this site and, therefore, to identify more candidates of sulfation. The specific goals are: 1. to identify the characteristics of sulfated tyrosines which distinguish them from non-sulfated tyrosines. These characteristics include the number and from non-sulfated tyrosines. These characteristics include the possible distribution of amino acids, as well as the possible secondary structural secondary structural features in the vicinity of the tyrosine. 2. to identify features in the vicinity of the tyrosine. 2. to identify proteins with probable proteins with probable tyrosine sulfation sites from the Swiss Protein data base. 3. to determine sequence similarities among the set of known proteins in tyrosine sulfation sites from the Swiss Protein data base. 3. to determine sequence similarities among the set of known proteins in the vicinity of the the vicinity of the sulfation site. Identification of new protein sulfation sites may be essential for the understanding of the function of these proteins sulfation site. Identification of new protein sulfation sites may be essential and will lead to a better understanding of the function of tyrosine sulfation. for the understanding of the function of these proteins and will lead to a better understanding of the function of tyrosine sulfation. In fact, approximately 1% of the tyrosines are sulfated in tissue in the rat. Despite this observation, only 24 unique protein sequences containing tyrosine sulfate have been described. The aim of this project is to analyze the amino acid sequences surrounding know tyrosine sulfation sites to determine the characteristics of this site and, therefore, to identify more candidates of sulfation. The specific goals are: 1. to identify the characteristics of sulfated tyrosines which distinguish them from non-sulfated tyrosines. These characteristics include the number and from non-sulfated tyrosines. These characteristics include the possible distribution of amino acids, as well as the possible secondary structural secondary structural features in the vicinity of the tyrosine. 2. to identify features in the vicinity of the tyrosine. 2. to identify proteins with probable proteins with probable tyrosine sulfation sites from the Swiss Protein data base. 3. to determine sequence similarities among the set of known proteins in tyrosine sulfation sites from the Swiss Protein data base. 3. to determine sequence similarities among the set of known proteins in the vicinity of the the vicinity of the sulfation site. Identification of new protein sulfation sites may be essential for the understanding of the function of these proteins sulfation site. Identification of new protein sulfation sites may be essential and will lead to a better understanding of the function of tyrosine sulfation. for the understanding of the function of these proteins and will lead to a better understanding of the function of tyrosine sulfation. Publications - Chan, Steve, S., Nicholas, Hugh B., Jr., and Rosenquist, Grace L.: "Re-evaluation of the Determinants of Tyrosine Sulfation": (In manuscript) Achievements - 1. Statistical analysis of the amino acids surrounding the target tyrosine using a specific scoring matrix (PSSM) showed that the consensus sequence does not contain the information to predict tyrosine sulfation. Previously, tyrosine sulfation had been thought to involve recognition of speciic features of a consensus sequence by the sulfating enzyme, tyrosyl-protein sulfotransferase (TPST). 2. Accurate prediction of tyrosine sulfation requires consideration of all residues within a window of 5 amino acids on either side of the tyrosine. 3. We conclude that highly specific recognition features are not required for substrate recognition by TPST. TPST may instead broadly recognize and act on any tyrosine residue sufficiently exposed to the solvent. Publications - Chan, Steve, S., Nicholas, Hugh B., Jr., and Rosenquist, Grace L.: "Re-evaluation of the Determinants of Tyrosine Sulfation": (In manuscript) Achievements - 1. Statistical analysis of the amino acids surrounding the target tyrosine using a specific scoring matrix (PSSM) showed that the consensus sequence does not contain the information to predict tyrosine sulfation. Previously, tyrosine sulfation had been thought to involve recognition of speciic features of a consensus sequence by the sulfating enzyme, tyrosyl-protein sulfotransferase (TPST). 2. Accurate prediction of tyrosine sulfation requires consideration of all residues within a window of 5 amino acids on either side of the tyrosine. 3. We conclude that highly specific recognition features are not required for substrate recognition by TPST. TPST may instead broadly recognize and act on any tyrosine residue sufficiently exposed to the solvent. Publications - Chan, Steve, S., Nicholas, Hugh B., Jr., and Rosenquist, Grace L.: "Re-evaluation of the Determinants of Tyrosine Sulfation": (In manuscript) Achievements - 1. Statistical analysis of the amino acids surrounding the target tyrosine using a specific scoring matrix (PSSM) showed that the consensus sequence does not contain the information to predict tyrosine sulfation. Previously, tyrosine sulfation had been thought to involve recognition of speciic features of a consensus sequence by the sulfating enzyme, tyrosyl-protein sulfotransferase (TPST). 2. Accurate prediction of tyrosine sulfation requires consideration of all residues within a window of 5 amino acids on either side of the tyrosine. 3. We conclude that highly specific recognition features are not required for substrate recognition by TPST. TPST may instead broadly recognize and act on any tyrosine residue sufficiently exposed to the solvent. Publications - Nicholas, Jr., H.B., Chan, S.S., Rosenquist, G.L.: "Reevaluation of the Determinants of Tyrosine Sulfation": Endocrine: 11:285-292: 1999.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 硫酸化是在所有含有高尔基体系统的动物细胞中发现的一种广泛的酪氨酸翻译后修饰。我们最近对酪氨酸硫酸化位点的分析表明，几乎一半包含酪氨酸运输基序，Y-x-x-疏水性（x 是任何氨基酸）。由于含有酪氨酸位点的蛋白质是在反高尔基体中加工的，那么是什么引导它们到达正确的目的地呢？我们将运输基序与酪氨酸硫酸化位点进行了对比，两者都包含 Y-x-x-疏水模式，以确定区分这些不同的基于酪氨酸的位点的序列特征。创建了几个对数比位置特定评分矩阵（PSSM）来区分硫酸化位点和运输基序。硫酸化位点和贩运主题相互对比，并与 Swiss-Prot 后生动物数据库进行对比。十三个贩运基序中的三个在已知的酪氨酸硫酸化位点范围内得分。 PSSM 的评估表明，高分氨基酸的模式因运输基序和硫酸化位点而异。在运输基序中，arg 和 gln 在位置 y+1 处优先，gly 在位置 y-1 处优先，而在硫酸化位点中，小残基（asp、gly、pro）在 y+1 处优先，而 asp 在 y-1 处优先。在运输基序中，y+3 处优选亮氨酸，而在硫酸化位点中优选蛋氨酸和色氨酸。这些结果表明，尽管酪氨酸硫酸化位点和运输基序之间存在显着差异，但这两个基于酪氨酸的位点之间存在一些重叠。进一步的实验应该确定一些运输基序是否在体内被硫酸化，以及硫酸化是否影响蛋白质的最终目的地。可能需要其他因素（例如其他基序）将蛋白质引导至正确的目的地。事实上，大鼠组织中大约 1% 的酪氨酸被硫酸化。尽管有这一观察结果，但仅描述了 24 个含有酪氨酸硫酸盐的独特蛋白质序列。该项目的目的是分析已知酪氨酸硫酸化位点周围的氨基酸序列，以确定该位点的特征，从而识别更多的硫酸化候选者。具体目标是： 1. 鉴定硫酸化酪氨酸与非硫酸化酪氨酸的区别特征。这些特征包括非硫酸化酪氨酸的数量和数量。这些特征包括氨基酸可能的分布，以及酪氨酸附近可能的二级结构特征。 2. 识别酪氨酸附近的特征。 2.从瑞士蛋白质数据库中鉴定具有可能酪氨酸硫酸化位点的可能蛋白质。 3. 确定瑞士蛋白质数据库中酪氨酸硫酸化位点的已知蛋白质组之间的序列相似性。 3.确定硫酸化位点附近的一组已知蛋白质之间的序列相似性。新蛋白质硫酸化位点的鉴定对于理解这些蛋白质硫酸化位点的功能可能至关重要。鉴定新的蛋白质硫酸化位点可能至关重要，并将有助于更好地了解酪氨酸硫酸化的功能。为了了解这些蛋白质的功能，并将导致更好地了解酪氨酸硫酸化的功能。事实上，大鼠组织中大约 1% 的酪氨酸被硫酸化。尽管有这一观察结果，但仅描述了 24 个含有酪氨酸硫酸盐的独特蛋白质序列。该项目的目的是分析已知酪氨酸硫酸化位点周围的氨基酸序列，以确定该位点的特征，从而识别更多的硫酸化候选者。具体目标是： 1. 鉴定硫酸化酪氨酸与非硫酸化酪氨酸的区别特征。这些特征包括非硫酸化酪氨酸的数量和数量。这些特征包括氨基酸可能的分布，以及酪氨酸附近可能的二级结构特征。 2. 识别酪氨酸附近的特征。 2.从瑞士蛋白质数据库中鉴定具有可能酪氨酸硫酸化位点的可能蛋白质。 3. 确定瑞士蛋白质数据库中酪氨酸硫酸化位点的已知蛋白质组之间的序列相似性。 3.确定硫酸化位点附近的一组已知蛋白质之间的序列相似性。新蛋白质硫酸化位点的鉴定对于理解这些蛋白质硫酸化位点的功能可能至关重要。鉴定新的蛋白质硫酸化位点可能至关重要，并将有助于更好地了解酪氨酸硫酸化的功能。为了了解这些蛋白质的功能，并将导致更好地了解酪氨酸硫酸化的功能。事实上，大鼠组织中大约 1% 的酪氨酸被硫酸化。尽管有这一观察结果，但仅描述了 24 个含有酪氨酸硫酸盐的独特蛋白质序列。该项目的目的是分析已知酪氨酸硫酸化位点周围的氨基酸序列，以确定该位点的特征，从而识别更多的硫酸化候选者。具体目标是： 1. 鉴定硫酸化酪氨酸与非硫酸化酪氨酸的区别特征。这些特征包括非硫酸化酪氨酸的数量和数量。这些特征包括氨基酸可能的分布，以及酪氨酸附近可能的二级结构特征。 2. 识别酪氨酸附近的特征。 2.从瑞士蛋白质数据库中鉴定具有可能酪氨酸硫酸化位点的可能蛋白质。 3. 确定瑞士蛋白质数据库中酪氨酸硫酸化位点的已知蛋白质组之间的序列相似性。 3.确定硫酸化位点附近的一组已知蛋白质之间的序列相似性。新蛋白质硫酸化位点的鉴定对于理解这些蛋白质硫酸化位点的功能可能至关重要。鉴定新的蛋白质硫酸化位点可能至关重要，并将有助于更好地了解酪氨酸硫酸化的功能。为了了解这些蛋白质的功能，并将导致更好地了解酪氨酸硫酸化的功能。出版物 - Chan, Steve, S., Nicholas, Hugh B., Jr. 和 Rosenquist, Grace L.：“重新评估酪氨酸硫酸化的决定因素”：（手稿中）成就 - 1. 使用特定评分矩阵 (PSSM) 对目标酪氨酸周围的氨基酸进行统计分析，表明共有序列不包含预测酪氨酸硫酸化的信息。此前，酪氨酸硫酸化被认为涉及硫酸化酶酪氨酰蛋白磺基转移酶（TPST）对共有序列的特定特征的识别。 2. 酪氨酸硫酸化的准确预测需要考虑酪氨酸两侧 5 个氨基酸窗口内的所有残基。 3. 我们的结论是，TPST 识别底物不需要高度特异性的识别特征。相反，TPST 可以广泛识别并作用于充分暴露于溶剂的任何酪氨酸残基。出版物 - Chan, Steve, S., Nicholas, Hugh B., Jr. 和 Rosenquist, Grace L.：“重新评估酪氨酸硫酸化的决定因素”：（手稿中）成就 - 1. 使用特定评分矩阵 (PSSM) 对目标酪氨酸周围的氨基酸进行统计分析，表明共有序列不包含预测酪氨酸硫酸化的信息。此前，酪氨酸硫酸化被认为涉及硫酸化酶酪氨酰蛋白磺基转移酶（TPST）对共有序列的特定特征的识别。 2. 酪氨酸硫酸化的准确预测需要考虑酪氨酸两侧 5 个氨基酸窗口内的所有残基。 3. 我们的结论是，TPST 识别底物不需要高度特异性的识别特征。相反，TPST 可以广泛识别并作用于充分暴露于溶剂的任何酪氨酸残基。出版物 - Chan, Steve, S., Nicholas, Hugh B., Jr. 和 Rosenquist, Grace L.：“重新评估酪氨酸硫酸化的决定因素”：（手稿中）成就 - 1. 使用特定评分矩阵 (PSSM) 对目标酪氨酸周围的氨基酸进行统计分析，表明共有序列不包含预测酪氨酸硫酸化的信息。此前，酪氨酸硫酸化被认为涉及硫酸化酶酪氨酰蛋白磺基转移酶（TPST）对共有序列的特定特征的识别。 2. 酪氨酸硫酸化的准确预测需要考虑酪氨酸两侧 5 个氨基酸窗口内的所有残基。 3. 我们的结论是，TPST 识别底物不需要高度特异性的识别特征。相反，TPST 可以广泛识别并作用于充分暴露于溶剂的任何酪氨酸残基。 出版物 - Nicholas, Jr., H.B., Chan, S.S., Rosenquist, G.L.：“酪氨酸硫酸化决定因素的重新评估”：内分泌：11：285-292：1999。