公开(公告)号：US5639869A

公开(公告)日：1997-06-17

申请号：US165038

申请日：1993-12-10

申请人： Barry C. Cole ,  Curtis L. Atkin ,  Arnold R. Oliphant ,  Ann Pole

发明人： Barry C. Cole ,  Curtis L. Atkin ,  Arnold R. Oliphant ,  Ann Pole

IPC分类号： A61K38/00 ,  C07H21/04 ,  C07K14/30

CPC分类号： C07H21/04 ,  C07K14/30 ,  A61K38/00 ,  Y10S530/825

摘要： A method of purifying Mycoplasma arthritidis mitogen (MAM) to electrophoretic and sequence homogeneity is disclosed. A preparation of MAM purified according to this method was used to determine the sequence of the N-terminal 54 amino acids of MAM. A synthetic peptide consisting of amino acids 15-32 inhibited MAM-induced cell proliferation in vitro. The sequence of the N-terminal 54 amino acids was reverse translated, nucleotide probes were designed therefrom, and the MAM gene was selected from a genomic library. The MAM gene was sequenced and found to be contained on a 1107 bp DNA fragment. The primary translation product contains a 39 amino acid signal sequence and a 213 amino acid mature MAM (molecular weight 25,294). Amino acid sequence comparisons of MAM to bacterial and murine tumor virus superantigens showed regions of conservative sequence homology, including the region capable of inhibiting cell proliferation. Sequence homologies to HIV and other retrovirus proteins and to certain regulatory proteins were also detected. Strategies for blocking or immunizing against certain diseases, including autoimmune diseases, are disclosed.

摘要翻译： 公开了一种纯化支原体支原体丝裂原（MAM）以电泳和序列同质性的方法。 使用根据该方法纯化的MAM制剂来确定MAM的N-末端54个氨基酸的序列。 由氨基酸15-32组成的合成肽在体外抑制MAM诱导的细胞增殖。 将N-末端54个氨基酸的序列反向翻译，设计核苷酸探针，并从基因组文库中选择MAM基因。 对MAM基因进行测序，发现其含有1107bp的DNA片段。 初级翻译产物含有39个氨基酸的信号序列和213个氨基酸的成熟MAM（分子量为25,294）。 MAM与细菌和鼠肿瘤病毒超级抗原的氨基酸序列比较显示出保守序列同源性的区域，包括能够抑制细胞增殖的区域。 还检测到与HIV和其他逆转录病毒蛋白质和某些调节蛋白质的序列同源性。 公开了阻止或免疫某些疾病（包括自身免疫性疾病）的策略。

公开(公告)号：US5872233A

公开(公告)日：1999-02-16

申请号：US876781

申请日：1997-06-17

申请人： Barry C. Cole ,  Curtis L. Atkin ,  Arnold R. Oliphant ,  Ann Pole

发明人： Barry C. Cole ,  Curtis L. Atkin ,  Arnold R. Oliphant ,  Ann Pole

IPC分类号： A61K38/00 ,  C07H21/04 ,  C07K14/30

CPC分类号： C07H21/04 ,  C07K14/30 ,  A61K38/00 ,  Y10S530/825

摘要： A method of purifying Mycoplasma arthritidis mitogen (MAM) to electrophoretic and sequence homogeneity is disclosed. A preparation of MAM purified according to this method was used to determine the sequence of the N-terminal 54 amino acids of MAM. A synthetic peptide consisting of amino acids 15-32 inhibited MAM-induced cell proliferation in vitro. The sequence of the N-terminal 54 amino acids was reverse translated, nucleotide probes were designed therefrom, and the MAM gene was selected from a genomic library. The MAM gene was sequenced and found to be contained on a 1107 bp DNA fragment. The primary translation product contains a 39 amino acid signal sequence and a 213 amino acid mature MAM (molecular weight 25,294). Amino acid sequence comparisons of MAM to bacterial and murine tumor virus superantigens showed regions of conservative sequence homology, including the region capable of inhibiting cell proliferation. Sequence homologies to HIV and other retrovirus proteins and to certain regulatory proteins were also detected. Strategies for blocking or immunizing against certain diseases, including autoimmune diseases, are disclosed.

摘要翻译： 公开了一种纯化支原体支原体丝裂原（MAM）以电泳和序列同质性的方法。 使用根据该方法纯化的MAM制剂来确定MAM的N-末端54个氨基酸的序列。 由氨基酸15-32组成的合成肽在体外抑制MAM诱导的细胞增殖。 将N-末端54个氨基酸的序列反向翻译，设计核苷酸探针，并从基因组文库中选择MAM基因。 对MAM基因进行测序，发现其含有1107bp的DNA片段。 初级翻译产物含有39个氨基酸的信号序列和213个氨基酸的成熟MAM（分子量为25,294）。 MAM与细菌和鼠肿瘤病毒超级抗原的氨基酸序列比较显示出保守序列同源性的区域，包括能够抑制细胞增殖的区域。 还检测到与HIV和其他逆转录病毒蛋白质和某些调节蛋白质的序列同源性。 公开了阻止或免疫某些疾病（包括自身免疫性疾病）的策略。

公开(公告)号：US5795974A

公开(公告)日：1998-08-18

申请号：US621081

申请日：1996-03-22

申请人： Barry C. Cole ,  Curtis L. Atkin ,  Kevin L. Knudtson

发明人： Barry C. Cole ,  Curtis L. Atkin ,  Kevin L. Knudtson

IPC分类号： A61K38/00 ,  C07H21/04 ,  C07K14/30

CPC分类号： C07K14/30 ,  C07H21/04 ,  A61K38/00

摘要： The gene encoding the superantigen Mycoplasma arthritidis T-cell mitogen (MAM) was molecularly cloned. The mam gene was modified by site-directed mutagenesis to change UGA codons, which in mycoplasmas are read as tryptophan codons instead of universal stop codons, to UGG codons such that the gene could be expressed in standard expression systems. Recombinant MAM, including extra amino acid residues at the N- and C-termini, were expressed and discovered to be biologically active. Certain amino acid substitutions in active regions of the protein also yield biologically active MAM proteins. A method of diagnosing rheumatoid arthritis using recombinant MAM protein in an ELISA assay is disclosed.

摘要翻译： 编码超抗原支原体支原体T细胞有丝分裂原（MAM）的基因被分子克隆。 通过定点诱变修饰mam基因，以将UGA密码子改变为UGA密码子，其将支原体作为色氨酸密码子而不是通用终止密码子读取到UGG密码子，使得该基因可以在标准表达系统中表达。 表达并发现重组MAM，包括N-和C-末端的额外的氨基酸残基是具有生物活性的。 蛋白质活性区域中的某些氨基酸取代也产生生物活性的MAM蛋白质。 公开了一种在ELISA测定中使用重组MAM蛋白诊断类风湿性关节炎的方法。