[发明专利]人类细胞中体的蛋白质复合物及其分离方法和用途

摘要

本发明涉及人体细胞分离技术。所述的人类细胞中体的蛋白质复合物，它至少由129种蛋白组成，其中有30种细胞骨架蛋白、4种中心体蛋白、6种囊泡转运相关蛋白、6种染色体调节蛋白、2种马达蛋白、3种检测点蛋白、7种降解调控蛋白、9种膜蛋白、12种转录与翻译相关蛋白、28种蛋白激酶、5种线粒体蛋白、9种分子伴侣蛋白和7种未知类蛋白。在分离方法中，包括将人体细胞培养同步化及收集、加入低渗液重悬并孵育使收集细胞膨胀、破碎重悬的膨胀细胞后取出上清、通过密度梯度进行离心分离并利用去污剂抽提纯化蛋白质复合物、分离鉴定中体蛋白种类。人类细胞中体的蛋白质复合物组成的揭示为筛选抗肿瘤药物提供了模式系统和靶标蛋白ezrin及靶位。

主权项

1.人类细胞中体的蛋白质复合物，其特征在于，它至少由129种蛋白组成，其具体蛋白种类名称及相关的国际数据库(NCBI)编号为： 序号 蛋白名称 数据库编号 1 Plectin 1 Q15149-2 2 Spectrin alpha chain P02549 3 Spectrin beta chain Q01082 4 Filamin A P21333 5 Filamin A O75369 6 abnormal spindle protein ASP Q8IUL2 7 Ankyrin 1 P16157 8 Nonmuscle myosin heavy chain-A P35579 9 Myosin Va Q9Y4I1 10 Myosin light chain kinase Q15746 11 Microtubule-actin crosslinking factor 1 Q9UPN3 12 Microtubule-associated protein 4 P27816 13 Microtubule-associated protein 2 P11137 14 Tubulin beta-5 chain P05218 15 Tubulin beta-1 chain P07437 16 Tubulin beta-2 chain P05217 17 Alpha-tubulin 1 P05209 18 Alpha-tubulin 6 Q9BQE3 19 Non-muscle alpha-actinin 4 O43707 20 Beta-actin P02570 21 Gamma-actin P02571 22 Vimentin P08670 23 Keratin，type II cytoskeletal 8 P05787 24 cytokeratin 18 P05783 25 Cytokeratin 19 P08727 26 Dynamin 3 Q9UQ16 27 Cytokeratin 17 Q04695 28 Protein Tara Q9H2D6 29 Nuclear envelope spectrin repeat protein 2 Q8WXH0 30 Huntingtin interacting protein 1 O00291 31 Tubulin gamma-1 chain P23258 32 gamma-tubulin complex protein 2 Q9BSJ2-2 33 nuclear mitotic apparatus protein 1 Q14981 34 Kendrin O95613 35 Cytoplasmic dynein heavy chain 1 Q14204 36 Dynactin complex 50kDa subunit Q13561 37 Dynactin 1 Q14203 38 Rab6-KIFL O95235 39 Golgin-245 Q13439 40 Nuclear RNA export factor 5 Q9H1B4 41 DNA topoisomerase II P11388 42 Serine/threonine protein kinase 12 Q96GD4 43 Double-strand-break repair protein rad21 homolog O60216 44 tumor protein p53 Q9HAQ8 45 Rad52 P43351 46 SWI/SNF-related matrix-associated actin-dependent regulator of chromatin d2 Q92925 47 CENP-E protein Q02224 48 Kinesin-related motor protein Eg5 P52732 49 hBUBR1 O60566 50 mitotic checkpoint Q13312 protein 1/Mad1 51 Cell division control protein 2 homolog P06493 52 suppressor of G2 allele of SKP1 Q9Y2Z0 53 26S protease regulatory subunit 4 Q03527 54 26S proteasome regulatory subunit S10 Q15008 55 SUMO-1-specific protease Q9GZR1-2 56 ubiquitin-specific protease Q9UHP3-2 57 Ubiquitin carboxyl-terminal hydrolase 7 Q93009 58 ubiquitin-conjugating enzyme E2 Q9Y2X8 59 Laminin gamma-1 chain precursor P11047 60 Lamin A/C P02545 61 Lamin B2 Q03252 62 Annexin II P07355 63 cadherin-associated protein Q9UQB3 64 Radixin P35241 65 Moesin P26038 66 ezrin P15311 67 Merlin P35240 68 Translation initiation factor IF-2 P46199 69 Eukaryotic translation initiation factor 2-alpha kinase 3 Q9NZJ5 70 Elongation factor 2 P13639 71 Elongation factor Tu P49411 72 Eukaryotic initiation factor 4A-I P04765 73 Retinoblastoma-like protein 2 Q08999 74 EBNA-2 co-activator Q13122 75 Thyroid hormone receptor-associated protein complex 100kDa component O75448 76 RuvB-like 2 Q9Y230 77 Protein disulfide isomerase A3 precursor P30101 78 chromatin-specific transcription elongation factor large subunit Q9Y5B9 79 Threonyl-tRNA synthetase P26639 80 Serine/threonine protein phosphatase 2A P05323 81 serine/threonine kinase 4(Ste20，yeast homolog) Q13043 82 Serine/threonine protein kinase 24 (STE20-like kinase MST3) Q9Y6E0 83 Serine/threonine protein kinase 9 O76039 84 protein tyrosine phosphatase Q12923-3 85 activated p21cdc42Hs kinase Q07912 86 cell division cycle 2-like 2 isoform 2 Q9UQ88-5 87 Tyrosine-protein kinase JAK1 P23458 88 protein tyrosine kinase FAK Q05397 89 M-phase phosphoprotein 9 Q9H976 90 RasGAP-activating-like protein 1 O95294 91 Rho GTPase activating protein 6 isoform 5 O43182-2 92 Ras P46940 GTPase-activating-like protein 93 Ras GTPase-activating protein 3 Q14644 94 Rho guanine nucleotide exchange factor 1 Q92888 95 Similar to SLIT-ROBO Rho GTPase-activating protein 1 Q8N6J5 96 Guanine nucleotide-binding protein，alpha-14 subunit O95837 97 Ras guanine nucleotide exchange factor son-of-sevenless(sos) 1 Q07889 98 Caspase recruitment domain protein 15 Q9HC29 99 mitogen-activated protein kinase 9 P45984-3 100 Mitogen-activated protein kinase 10 P53779 101 IQGAP1 P46940 102 Citron protein O14578 103 Transketolase P29401 104 Alpha enolase P06733 105 Centaurin delta 2 Q96P48 106 Helicase-like protein Q9NZ71 107 heterogeneous nuclear ribonucleoprotein A2/B1 P22626-2 108 Creatine kinase， ubiquitous mitochondrial precursor P12532 109 Pyruvate kinase，M1 isozyme P14618 110 Pyruvate carboxylase， mitochondrial precursor P11498 111 Carbamoyl-phosphate synthase P31327 112 Ubiquinol-cytochrome C reductase complex core protein 2 P22695 113 Heat shock cognate 71 kDa protein P11142 114 Stress-70 protein， mitochondrial precursor P38646 115 Heat shock 70kDa protein 1(HSP70.1) P08107 116 Heat shock protein 75 kDa，mitochondrial precursor(HSP 75) Q12931 117 ATP synthase beta chain，mitochondrial precursor P06576/P2 5705 118 ATP-dependent RNA helicase A Q08211 119 T-complex protein 1 P50990 120 78kDa glucose-regulated protein precursor(GRP 78) P11021 121 XPA-binding protein 2 Q9HCS7 122 dJ337018.7(novel protein) Q96MP5 123 Dolichyl-diphosphoolig osaccharide--protein glycosyltransferase 67 kDa subunit precursor P04843 124 ubiquitin UBF-fl Q96SQ5 125 26S proteasome non-ATPase regulatory subunit 12 O00232 126 similar to KIAA0342 XP047357 127 KIAA0465 BAA32310 128 mannose receptor Ctype 1 precursor P22897 129 Similar to early Q8IYZ2 hematopoietic zinc finger