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公开(公告)号：US07973132B2
公开(公告)日：2011-07-05
申请号：US12245546
申请日：2008-10-03
申请人： Ronald T. Raines , Stephen M. Fuchs
发明人： Ronald T. Raines , Stephen M. Fuchs
CPC分类号： C12Q1/37 , C07K14/43595 , G01N33/5005
摘要： This invention relates to methods and compositions for designing novel fluorescent proteins, preferably to a green fluorescent proteins (GFP). The engineered GFPs are modified by substituting negatively charged amino acids with positively charged amino acids on the exterior of the protein making the protein cell permeable. The ability of the engineered fluorescent proteins to permeate cells obviates the need for transfections, allowing these novel proteins to be used in numerous biological applications.
摘要翻译： 本发明涉及用于设计新的荧光蛋白，优选绿色荧光蛋白（GFP）的方法和组合物。 通过在蛋白质外部用带正电荷的氨基酸取代带负电荷的氨基酸来修饰工程化的GFP，使得蛋白质细胞是可渗透的。 工程化荧光蛋白渗透细胞的能力消除了转染的需要，允许这些新型蛋白质用于许多生物应用。
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公开(公告)号：US07655757B2
公开(公告)日：2010-02-02
申请号：US11454418
申请日：2006-06-16
摘要： This invention relates to cytotoxic variants of human ribonuclease 1 (RNase 1) identified through analysis of the interaction between RNase 1 and the human ribonuclease inhibitor (hRI) as defined by the three dimensional (3-D) atomic structure of the RNase 1 hRI complex. Also disclosed is the 3-D structure of the hRI.RNase 1 complex and methods for designing the RNase 1 variants.
摘要翻译： 本发明涉及通过分析核糖核酸酶1和核糖核酸酶抑制剂（hRI）之间的相互作用鉴定的人核糖核酸酶1（RNase 1）的细胞毒性变体，如RNase 1 hRI复合物的三维（3-D）原子结构所定义 。 还公开了hRI.RNase 1复合物的3-D结构和用于设计RNA酶1变体的方法。
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公开(公告)号：US07560248B1
公开(公告)日：2009-07-14
申请号：US09234028
申请日：1999-01-20
申请人： Ronald T. Raines
发明人： Ronald T. Raines
CPC分类号： C07K14/4703
摘要： Mutant forms of ribonuclease inhibitor are described which are rendered more resistant to oxidation while retaining affinity for both ribonuclease and angiogenin. The mutant forms have another amino acid, typically an alanine, substituted for one or more of the adjacent cysteine residues in the wild-type sequence to prevent the formation of unwanted disulfide bonds which can disrupt the effectiveness of the molecule.
摘要翻译： 描述了核糖核酸酶抑制剂的突变形式，其对氧化具有更强的抗性，同时保持对核糖核酸酶和血管生成素的亲和力。 突变体形式具有另一个氨基酸，通常为丙氨酸，其取代了野生型序列中一个或多个相邻的半胱氨酸残基，以防止可能破坏分子有效性的不想要的二硫键。
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公开(公告)号：US07098016B2
公开(公告)日：2006-08-29
申请号：US10461713
申请日：2003-06-13
IPC分类号： C12N9/22
CPC分类号： C12N9/22
摘要： An enzyme is re-engineered to be a zymogen, an enzyme precursor which is converted into an enzyme by protease cleavage. In the example described here, an RNase A enzyme is converted into a zymogen by adding to the enzyme a bridge of amino acids linking the amino and carboxyl termini of the enzyme. The bridge has built in it a protease cleavage site for a specific protease, for example the protease plasmepsin II, produced by the malaria parasite. Since RNase A can be made cytotoxic, this permits a cytotoxic enzyme to be made in the form of a zymogen that becomes active only when it is acted on by a protease only present in a particular target cell such as a pathogen.
摘要翻译： 酶被重新设计为酶原，酶原，其通过蛋白酶切割转化成酶。 在本文所述的实施例中，通过向酶加入连接酶的氨基和羧基末端的氨基酸桥，将RNase A酶转化为酵素。 该桥已经建立了特定蛋白酶的蛋白酶切割位点，例如由疟原虫产生的蛋白酶等质粒蛋白酶II。 由于RNA酶A可以被制成细胞毒性，因此允许细胞毒素酶以酶原的形式制备，只有当其被存在于特定靶细胞例如病原体中的蛋白酶作用时才变成活性。
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公开(公告)号：US5840296A
公开(公告)日：1998-11-24
申请号：US950866
申请日：1997-10-15
摘要： Modified ribonucleases belonging to the RNase A superfamily of ribonucleases is disclosed. Each modified ribonuclease has a mutation in the loop region corresponding to amino acids 85-94 of bovine pancreatic RNase A. Each modified ribonuclease has reduced binding affinity for ribonuclease inhibitor (RI), wild-type ribonuclease activity, and exhibits enhanced cytotoxicity toward tumor cells, relative to the wild-type ribonuclease. Also disclosed is a method for obtaining a modified ribonuclease having reduced binding affinity for RI, wild-type ribonuclease activity, and enhanced cytotoxicity.
摘要翻译： 公开了属于核糖核酸酶A核糖核酸酶超家族的修饰的核糖核酸酶。 每个修饰的核糖核酸酶在对应于牛胰腺核糖核酸酶A的氨基酸85-94的环区域中具有突变。每个修饰的核糖核酸酶具有降低的核糖核酸酶抑制剂（RI）的结合亲和力，野生型核糖核酸酶活性，并且表现出对肿瘤细胞的增强的细胞毒性 ，相对于野生型核糖核酸酶。 还公开了获得具有降低的RI的结合亲和力，野生型核糖核酸酶活性和增强的细胞毒性的修饰的核糖核酸酶的方法。
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