Peptide Bond Synthesis at Monolayer Film Interfaces

单层膜界面的肽键合成

• 批准号: 9508066
• 负责人: John Oliver
• 金额: $ 4.2万
• 依托单位: Brown University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-06-01 至 1997-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9508066&HistoricalAwards=false
• 关键词: Peptide; Bond; Synthesis; Monolayer; Film

; R o o t E n t r y F C o m p O b j b W o r d D o c u m e n t O b j e c t P o o l 6 7 8 9 : ; = ? F Microsoft Word 6.0 Document MSWordDoc Word.Document.6 ; @ @ @ p @ Oliver 9508066 The primary objective of this research is to demonstrate that peptides can be synthesized from suitably derivatized and aminoacylated nucleotides which are suspended as a monolayer film at a water/air interface. Successful peptide synthesis at a monolayer surface may be a good mimic for ribosomal peptide synthesis as well as an efficient method to synthesize certain peptides. The formation of amide bonds from ester bonds is energetically downhill. However, biological systems find it necessary to catalyze peptide synthesis from activated amino acids with proteins or protein/RNA complexes. The catalysts perform at least two functions. The first role is to accelerate amide bond formation so that it competes with non-enzymatic hydrolysis of the activated amino acids. In addition the catalyst imposes order on the synthesis so that only a defined polymer is produced. We have now demonstrated that lipid derivatized nucleotides form monolayer films and these films can be compressed to densities similar to those seen in solids. This provides a compelling motivation to investigate the possibi lity of synthesizing peptide bonds at the monolayer. Nucleotides conjugated to lipids at the 5'-phosphate and aminoacylated at the 2',3'-position will be used to form a monolayer film. The surface pressure and packing density in the monolayer can be controlled by physically changing the surface area of the interface. At high packing densities we expect to see amide bond fomlation with rates higher than background hydrolysis. It is expected that the results obtained from this research will be both general and important, with significant implications for enzyme mechanisms and chemical peptide synthesis. %%% The goal of this award is to demonstrate that polypeptides can be synthesized in a non-enzymatic system at rates approaching those seen with ribosomes. We intend to synthesize derivatives of amino acids which will incorporate physical characteristics required to lock them at an air-water interface. Reaction rates at this interface will be controlled by varying the surface pressure andl thus the density of the amino acid derivatives. Infrared spectroscopy will be used to monitor the progress of the reaction. These preliminary experiments coupled with methods to preorganize the amino acid derivatives may allow us to design a method of rapidly synthesizing polypeptides of defined sequence. *** @ @ ; Oh +' 0 $ H l D h S u m m a r y I n f o r m a t i o n ( 5 R:\WWUSER\TEMPLATE\NORMAL.DOT jack cohen jack cohen @ { R @ G @ ug @ u Microsoft Word 6.0 7 e = e j j j j j j j g = ) I T ( g j g j j j j ~ j j j j a Oliver 9508066 The primary objective of this research is to demonstrate that peptides can be synthesized from suitably derivatized and aminoacylated nucleotides which are suspended as a monolayer film at a water/air interface. Successful peptide synthesis at a monolayer surface may be a good mimic for ribosomal peptide synthesis as well as an efficient method to synthesize certain peptides. The formation of amide bonds from ester bonds is energetically downhill. However, biological systems find it necessary to catalyze peptide synthesis from activated amino acids wit

;​R o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o6 7 8 9:；= ?F Microsoft Word 6.0文档MSWordDoc。6;​本研究的主要目的是证明肽可以由适当衍生化和氨基酰化的核苷酸合成，这些核苷酸作为单层膜悬浮在水/空气界面上。在单层表面成功合成多肽可能是核糖体合成多肽的良好模拟物，也是合成某些多肽的有效方法。由酯键形成酰胺键在能量上是下坡的。然而，生物系统发现有必要用蛋白质或蛋白质/RNA复合物催化活性氨基酸合成肽。催化剂至少有两种功能。第一个作用是加速酰胺键的形成，使其与非酶水解的活化氨基酸竞争。此外，催化剂对合成施加了秩序，因此只产生特定的聚合物。我们现在已经证明，脂质衍生的核苷酸形成单层膜，这些膜可以被压缩到与固体相似的密度。这为研究在单层上合成肽键的可能性提供了一个令人信服的动机。在5'-磷酸位点与脂质结合的核苷酸和在2‘,3’位点的氨基酰化的核苷酸将被用来形成单层膜。可以通过物理改变界面的表面积来控制单层中的表面压力和填充密度。在高堆积密度下，我们期望看到酰胺键形成的速率高于背景水解。预计本研究的结果将具有普遍性和重要性，对酶机制和化学肽合成具有重要意义。这个奖项的目标是证明多肽可以在非酶系统中以接近核糖体的速率合成。我们打算合成氨基酸的衍生物，它将包含将它们锁定在空气-水界面所需的物理特性。通过改变表面压力，从而改变氨基酸衍生物的密度，可以控制该界面的反应速率。红外光谱将用于监测反应的进展。这些初步实验与预先组织氨基酸衍生物的方法相结合，可以使我们设计一种快速合成确定序列的多肽的方法。*** @ @；Oh + 0 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $R:模板\ WWUSER \ \正常。DOT杰克科恩杰克科恩@ {R @ G @ ug @ u微软Word 6.0 7 e = ej j j j j j j j j g =) I T (g j g j j j j j本研究的主要目的是证明由适当的衍生化和氨基酰化的核苷酸在水/空气界面上作为单层膜悬浮可以合成肽。在单层表面成功合成多肽可能是核糖体合成多肽的良好模拟物，也是合成某些多肽的有效方法。由酯键形成酰胺键在能量上是下坡的。然而，生物系统发现有必要用活性氨基酸催化合成肽