公开(公告)号：US06864363B2

公开(公告)日：2005-03-08

申请号：US10008355

申请日：2001-11-08

申请人： James Travis ,  Jan S. Potempa ,  Agnieszka Banbula ,  Marcin Bugno

发明人： James Travis ,  Jan S. Potempa ,  Agnieszka Banbula ,  Marcin Bugno

IPC分类号： A61K38/00 ,  A61K39/00 ,  C12N9/48 ,  C07H21/04

CPC分类号： C12N9/48 ,  A61K38/00 ,  A61K39/00

摘要： The present invention provides isolated polypeptides, dipeptidylpeptidases, active analogs, active fragments, or active modifications thereof, having amidolytic activity for cleavage of a peptide bond between the second and third amino acids from the N-terminal end of a target polypeptide, wherein the target polypeptide has an aliphatic or an aromatic residue as a substituent on the α-carbon atom of the second amino acid from the N-terminal end of the peptide. Isolated nucleic acids encoding dipeptidylpeptidases are also provided, as are methods of reducing growth of a bacterium by inhibiting a dipeptidylpeptidase.

摘要翻译： 本发明提供分离的多肽，二肽基肽酶，活性类似物，活性片段或其活性修饰，其具有用于从目标多肽的N末端切割第二和第三个氨基酸之间的肽键的酰氨基活性，其中靶 多肽在肽的N末端的第二个氨基酸的α-碳原子上具有作为取代基的脂肪族或芳香族残基。 还提供了编码二肽基肽酶的分离的核酸，以及通过抑制二肽基肽酶来减少细菌生长的方法。

公开(公告)号：US06833262B1

公开(公告)日：2004-12-21

申请号：US10030330

申请日：2001-10-19

申请人： James Travis ,  Jan S. Potempa ,  Daniel C. Nelson

发明人： James Travis ,  Jan S. Potempa ,  Daniel C. Nelson

IPC分类号： C12N952

CPC分类号： C12N9/6472 ,  A61K39/00

摘要： An isolated oral bacterial polypeptide having amidolytic activity for cleavage of denatured polypeptides and nondenatured serpin polypeptides and particularly a human &agr;1-proteinase inhibitor polypeptide is provided. The mature polypeptide of the invention has a molecular weight of about 70 kD to about 80 kD. Also provided is an isolated nucleic acid sequence encoding the oral bacterial polypeptide of the invention, methods for identifying inhibitors of the polypeptide and compositions such as immunogenic compositions and inhibitor compositions.

摘要翻译： 提供了具有用于切割变性多肽和非变性丝氨酸蛋白酶多肽，特别是人α1-蛋白酶抑制剂多肽的酰胺分解活性的分离的口腔细菌多肽。 本发明的成熟多肽具有约70kD至约80kD的分子量。 还提供了编码本发明的口腔细菌多肽的分离的核酸序列，用于鉴定多肽的抑制剂的方法和组合物，例如免疫原性组合物和抑制剂组合物。

公开(公告)号：US5523390A

公开(公告)日：1996-06-04

申请号：US119361

申请日：1993-09-10

申请人： James Travis ,  Jan S. Potempa ,  Philip J. Barr ,  Nadine Pavloff

发明人： James Travis ,  Jan S. Potempa ,  Philip J. Barr ,  Nadine Pavloff

IPC分类号： C12N9/52 ,  C12N9/14 ,  C07H19/00 ,  C12N9/48

CPC分类号： C12N9/52

摘要： Provide herein is a substantially pure gingipain-1 preparation, gingipain-1 being characterized as having an apparent molecular mass of 50 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and an apparent molecular mass of 44 kDa as estimated by gel filtration chromatography, said gingipain-1 having amidolytic and proteolytic activity for cleavage after arginine residues and having no amidolytic and/or proteolytic activity for cleavage after lysine residues, wherein the amidolytic and/or proteolytic activity is inhibited by cysteine protease group-specific inhibitors including iodoacetamide, iodoacetic acid, N-ethylmaleimide, leupeptin, antipain, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, TLCK, TPCK, p-aminobenzamidine, N-chlorosuccinamide, and chelating agents including EDTA and EGTA, wherein the amidolytic and/or proteolytic activity of said gingipain-1 is not sensitive to inhibition by human cystatin C, .alpha.2-macroglobulin, .alpha.1-proteinase inhibitor, antithrombin III, .alpha.2-antiplasmin, serine protease group-specific inhibitors including diisopropylfluorophosphate, phenylmethyl sulfonylfluoride and 3,4-diisochlorocoumarin, and wherein the amidolytic and/or proteolytic activities of gingipain-1 are stabilized by Ca.sup.2+ and wherein the amidolytic and/or proteolytic activities of said gingipain-1 are stimulated by glycine-containing peptides and glycine analogues, and methods for preparation. As specifically exemplified, Arg-gingipain-1 and Arg-gingipain-2 are purified from Porphyromonas gingivalis, and the mature Arg-gingipain-2 has an amino acid sequence as given in SEQ ID NO:5 from amino acid 1 through amino acid 510. Also provided are nucleic acid sequences encoding Arg-gingipain-2. The nucleotide coding sequence of the mature Arg-gingipain-2 is given in SEQ ID NO:4, from nucleotide 1630 through nucleotide 3105.

摘要翻译： 本文提供的是基本上纯的姜黄素-1制剂，姜黄素-1的特征在于通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳估计的表观分子量为50kDa，通过凝胶过滤色谱法估计的表观分子量为44kDa，所述 姜黄素-1具有酰胺分解和蛋白水解活性，用于精氨酸残基之后的切割，并且在赖氨酸残基之后不具有用于切割的酰氨基裂解和/或蛋白水解活性，其中酰胺分解和/或蛋白水解活性被半胱氨酸蛋白酶组特异性抑制剂抑制，包括碘乙酰胺，碘乙酸 ，N-乙基马来酰亚胺，亮抑酶肽，抗胰蛋白酶，反式 - 环氧琥珀酰基-L-亮氨酰氨基 - （4-胍基）丁烷，TLCK，TPCK，对氨基苯甲脒，N-氯代琥珀酰胺和包括EDTA和EGTA的螯合剂，其中酰胺分解和/ 所述姜黄素-1的蛋白水解活性对人半胱氨酸蛋白酶抑制剂C，α2-巨球蛋白，α1-蛋白酶 e抑制剂，抗凝血酶III，α2-抗血纤维蛋白酶，丝氨酸蛋白酶组特异性抑制剂，包括二异丙基氟磷酸酯，苯基甲基磺酰氟和3,4-二氯代香豆素，其中盖蛋白-1的酰胺分解和/或蛋白水解活性由Ca 2+稳定，其中酰胺分解 和/或所述鹅蛋白-1的蛋白水解活性由含甘氨酸的肽和甘氨酸类似物及其制备方法刺激。 如具体示例，从牙龈卟啉单胞菌中纯化Arg-gingipain-1和Arg-gingipain-2，并且成熟的Arg-gingipain-2具有如SEQ ID NO：5中从氨基酸1至氨基酸510所给出的氨基酸序列 还提供了编码Arg-gingipain-2的核酸序列。 成熟Arg-gingipain-2的核苷酸编码序列在SEQ ID NO：4中由核苷酸1630至核苷酸3105给出。