公开(公告)号：US5196352A

公开(公告)日：1993-03-23

申请号：US722066

申请日：1991-06-27

申请人： John J. Siekierka ,  Hsuen-Yun Hung ,  Marie J. Staruch ,  Nolan H. Sigal ,  Richard A. Mumford

发明人： John J. Siekierka ,  Hsuen-Yun Hung ,  Marie J. Staruch ,  Nolan H. Sigal ,  Richard A. Mumford

IPC分类号： C12N9/90 ,  C12N15/90

CPC分类号： C12N15/90 ,  C12N9/90

摘要： A new homogeneous cytosolic binding (HCB) protein, having a specific binding activity of about 26 .mu.g FK-506 per mg protein and a molecular weight of about 10-12 kilodaltons, reversibly binds the immunosuppressant FK-506 but not cyclosporine A (CSA). The protein is stable to heating at 56 degrees C. for 30 minutes retaining its FK-506 binding affinity, and has the (partial) amino terminal amino acid sequence: H.sub.2 N G y V l G n V l G u T r I e S r P o G y A p G y A g T r P e P o L s A g-Gly-Gln-Thr-X-Val-Val-Val-His-Tyr-Thr-Gly-Met-Leu-Glu-Asp-Fly-Lys-Phe-ASp (wherein X is undefined). The HCBV protein is isolated from the cytosol of mammalian tissues, preferably human neoplastic T-cell lines, e.g. Jurkat, and can be used in diagnostic and purification procedures involving FK-506 macrolide type immunosuppressants. The HCB protein also catalyses the cis-trans isomerization of proline-containing peptide bonds.

摘要翻译： 具有约26μgFK-506 / mg蛋白质和约10-12千道尔顿分子量的特异结合活性的新的均匀胞质结合蛋白（HCB）可逆地结合免疫抑制剂FK-506而不是环孢菌素A（CSA ）。 蛋白质在56摄氏度下加热稳定30分钟，保持其FK-506结合亲和力，并具有（部分）氨基末端氨基酸序列：H2N G y V 1 G n V 1 G u T r I e S 其中，G-Gly-Gln-Thr-X-Val-Val-Val-His-Tyr-Thr-Gly-Met-Leu-Glu- Asp-Fly-Lys-Ph e-ASp（其中X未定义）。 HCBV蛋白从哺乳动物组织的细胞溶质中分离，优选人肿瘤T细胞系，例如， Jurkat，可用于涉及FK-506大环内酯型免疫抑制剂的诊断和纯化程序。 HCB蛋白还催化含脯氨酸肽键的顺式 - 反式异构化。

公开(公告)号：US5109112A

公开(公告)日：1992-04-28

申请号：US548429

申请日：1990-07-05

申请人： John J. Siekierka ,  Hsuen-Yun Hung ,  Marie J. Staruch ,  Nolan H. Sigal ,  Richard A. Mumford

发明人： John J. Siekierka ,  Hsuen-Yun Hung ,  Marie J. Staruch ,  Nolan H. Sigal ,  Richard A. Mumford

IPC分类号： C12N9/90 ,  C12N15/90

CPC分类号： C12N15/90 ,  C12N9/90 ,  Y10S530/81 ,  Y10S530/827

摘要： A new homogeneous cytosolic binding (HCB) protein, having a specific binding activity of about 26 .mu.g FK-506 per mg protein and a molecular weight of about 10-12 kilodaltons, reversibly binds the immunosuppressant FK-506 but not cyclosporine A (CSA). The protein is stable to heating at 56 degrees C. for 30 minutes retaining its FK-506 binding affinity, and has the (partial) amino terminal amino acid sequence: H.sub.2 N-Gly-Val-Gln-Val-Glu-Thr-Ile-Ser-Pro- Gly-Asp-Gly-Arg-Thr-Phe-Pro-Lys- Ar g-Gly-Gln-Thr-X-Val-Val-His-Tyr-Thr-Gly-Met-Leu-Glu-Asp-Gly-Lys-Lys-Phe-Asp (wherein X is undefined). The HCB protein is isolated from the cytosol of mammalian tissues, preferably human neoplastic T-cell lines, e.g., Jurkat, and can be used in diagnostic and purification procedures involving FK-506 macrolide type immunosuppressants. The HCB protein also catalyzes the cis-trans isomerization of proline-containing peptide bonds.

摘要翻译： 具有约26μgFK-506 / mg蛋白的特异性结合活性和约10-12千道尔顿分子量的新的均匀胞质结合（HCB）蛋白可逆地结合免疫抑制剂FK-506而不是环孢菌素A（CSA ）。 蛋白质在56℃下加热稳定30分钟，保持其FK-506结合亲和力，并具有（部分）氨基末端氨基酸序列：H2N-Gly-Val-Gln-Val-Glu-Thr-Ile- Ser-Pro-Gly-Asp-Gly-Arg-Thr-Phe-Pro-Lys-Ar g-Gly-Gln-Thr-X-Val-Val-His-Tyr-Thr-Gly-Met-Leu-Glu-Asp -Gly-Lys-Lys-Ph e-Asp（其中X未定义）。 HCB蛋白从哺乳动物组织的细胞溶质中分离，优选人类肿瘤T细胞系，例如Jurkat，并且可用于涉及FK-506大环内酯型免疫抑制剂的诊断和纯化方法。 HCB蛋白还催化含脯氨酸的肽键的顺式 - 反式异构化。