CN110092838B - 一种增强蛋白降解多肽的发现及应用 - Google Patents
一种增强蛋白降解多肽的发现及应用 Download PDFInfo
- Publication number
- CN110092838B CN110092838B CN201910383227.1A CN201910383227A CN110092838B CN 110092838 B CN110092838 B CN 110092838B CN 201910383227 A CN201910383227 A CN 201910383227A CN 110092838 B CN110092838 B CN 110092838B
- Authority
- CN
- China
- Prior art keywords
- protein
- polypeptide
- degradation
- sequence
- enhancing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 29
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 29
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 29
- 230000017854 proteolysis Effects 0.000 title claims abstract description 11
- 230000002708 enhancing effect Effects 0.000 title claims abstract description 9
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 25
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 13
- 238000006731 degradation reaction Methods 0.000 claims abstract description 12
- 230000015556 catabolic process Effects 0.000 claims abstract description 10
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 4
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 4
- 230000001105 regulatory effect Effects 0.000 claims abstract description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 230000001276 controlling effect Effects 0.000 claims description 3
- 210000003527 eukaryotic cell Anatomy 0.000 claims 1
- 201000010099 disease Diseases 0.000 abstract description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 230000004927 fusion Effects 0.000 abstract description 4
- 230000004845 protein aggregation Effects 0.000 abstract description 4
- 238000000338 in vitro Methods 0.000 abstract description 3
- 238000001727 in vivo Methods 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 abstract 1
- 230000015572 biosynthetic process Effects 0.000 abstract 1
- 239000002299 complementary DNA Substances 0.000 abstract 1
- 230000007281 self degradation Effects 0.000 abstract 1
- 238000003786 synthesis reaction Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 8
- 108010054624 red fluorescent protein Proteins 0.000 description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- 238000001890 transfection Methods 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 241000699660 Mus musculus Species 0.000 description 3
- 102000044159 Ubiquitin Human genes 0.000 description 3
- 108090000848 Ubiquitin Proteins 0.000 description 3
- 108091006047 fluorescent proteins Proteins 0.000 description 3
- 102000034287 fluorescent proteins Human genes 0.000 description 3
- 230000008685 targeting Effects 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 2
- 229930182816 L-glutamine Natural products 0.000 description 2
- 108010069926 arginyl-glycyl-serine Proteins 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 230000004770 neurodegeneration Effects 0.000 description 2
- 208000015122 neurodegenerative disease Diseases 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 238000003151 transfection method Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- FCXAUASCMJOFEY-NDKCEZKHSA-N Ala-Leu-Thr-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(O)=O FCXAUASCMJOFEY-NDKCEZKHSA-N 0.000 description 1
- MFAMTAVAFBPXDC-LPEHRKFASA-N Arg-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O MFAMTAVAFBPXDC-LPEHRKFASA-N 0.000 description 1
- XWSIYTYNLKCLJB-CIUDSAMLSA-N Asp-Lys-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O XWSIYTYNLKCLJB-CIUDSAMLSA-N 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- SBHVGKBYOQKAEA-SDDRHHMPSA-N Gln-His-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CCC(=O)N)N)C(=O)O SBHVGKBYOQKAEA-SDDRHHMPSA-N 0.000 description 1
- BIHMNDPWRUROFZ-JYJNAYRXSA-N Glu-His-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O BIHMNDPWRUROFZ-JYJNAYRXSA-N 0.000 description 1
- BRFJMRSRMOMIMU-WHFBIAKZSA-N Gly-Ala-Asn Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O BRFJMRSRMOMIMU-WHFBIAKZSA-N 0.000 description 1
- ZZJVYSAQQMDIRD-UWVGGRQHSA-N Gly-Pro-His Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O ZZJVYSAQQMDIRD-UWVGGRQHSA-N 0.000 description 1
- IRJWAYCXIYUHQE-WHFBIAKZSA-N Gly-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)CN IRJWAYCXIYUHQE-WHFBIAKZSA-N 0.000 description 1
- LCRDMSSAKLTKBU-ZDLURKLDSA-N Gly-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)CN LCRDMSSAKLTKBU-ZDLURKLDSA-N 0.000 description 1
- FIMNVXRZGUAGBI-AVGNSLFASA-N His-Glu-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O FIMNVXRZGUAGBI-AVGNSLFASA-N 0.000 description 1
- JBSLJUPMTYLLFH-MELADBBJSA-N His-His-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC3=CN=CN3)N)C(=O)O JBSLJUPMTYLLFH-MELADBBJSA-N 0.000 description 1
- ORERHHPZDDEMSC-VGDYDELISA-N His-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N ORERHHPZDDEMSC-VGDYDELISA-N 0.000 description 1
- FLXCRBXJRJSDHX-AVGNSLFASA-N His-Pro-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O FLXCRBXJRJSDHX-AVGNSLFASA-N 0.000 description 1
- VIJMRAIWYWRXSR-CIUDSAMLSA-N His-Ser-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 VIJMRAIWYWRXSR-CIUDSAMLSA-N 0.000 description 1
- CZCSUZMIRKFFFA-CIUDSAMLSA-N Leu-Ala-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O CZCSUZMIRKFFFA-CIUDSAMLSA-N 0.000 description 1
- ZFNLIDNJUWNIJL-WDCWCFNPSA-N Leu-Glu-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZFNLIDNJUWNIJL-WDCWCFNPSA-N 0.000 description 1
- QJXHMYMRGDOHRU-NHCYSSNCSA-N Leu-Ile-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O QJXHMYMRGDOHRU-NHCYSSNCSA-N 0.000 description 1
- SEMUSFOBZGKBGW-YTFOTSKYSA-N Leu-Ile-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O SEMUSFOBZGKBGW-YTFOTSKYSA-N 0.000 description 1
- QBEPTBMRQALPEV-MNXVOIDGSA-N Lys-Ile-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCCCN QBEPTBMRQALPEV-MNXVOIDGSA-N 0.000 description 1
- 208000001145 Metabolic Syndrome Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- UNBFGVQVQGXXCK-KKUMJFAQSA-N Phe-Ser-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O UNBFGVQVQGXXCK-KKUMJFAQSA-N 0.000 description 1
- AFXCXDQNRXTSBD-FJXKBIBVSA-N Pro-Gly-Thr Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O AFXCXDQNRXTSBD-FJXKBIBVSA-N 0.000 description 1
- WHNJMTHJGCEKGA-ULQDDVLXSA-N Pro-Phe-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O WHNJMTHJGCEKGA-ULQDDVLXSA-N 0.000 description 1
- PKHDJFHFMGQMPS-RCWTZXSCSA-N Pro-Thr-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O PKHDJFHFMGQMPS-RCWTZXSCSA-N 0.000 description 1
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 1
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 1
- HQTKVSCNCDLXSX-BQBZGAKWSA-N Ser-Arg-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O HQTKVSCNCDLXSX-BQBZGAKWSA-N 0.000 description 1
- GZBKRJVCRMZAST-XKBZYTNZSA-N Ser-Glu-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZBKRJVCRMZAST-XKBZYTNZSA-N 0.000 description 1
- QBUWQRKEHJXTOP-DCAQKATOSA-N Ser-His-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QBUWQRKEHJXTOP-DCAQKATOSA-N 0.000 description 1
- ASJDFGOPDCVXTG-KATARQTJSA-N Thr-Cys-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(O)=O ASJDFGOPDCVXTG-KATARQTJSA-N 0.000 description 1
- XSEPSRUDSPHMPX-KATARQTJSA-N Thr-Lys-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O XSEPSRUDSPHMPX-KATARQTJSA-N 0.000 description 1
- BPGDJSUFQKWUBK-KJEVXHAQSA-N Thr-Val-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 BPGDJSUFQKWUBK-KJEVXHAQSA-N 0.000 description 1
- 201000000690 abdominal obesity-metabolic syndrome Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 230000008236 biological pathway Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- TWFZGCMQGLPBSX-UHFFFAOYSA-N carbendazim Chemical compound C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 description 1
- 238000010523 cascade reaction Methods 0.000 description 1
- 230000006652 catabolic pathway Effects 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000008482 dysregulation Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 238000013537 high throughput screening Methods 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 210000003712 lysosome Anatomy 0.000 description 1
- 230000001868 lysosomic effect Effects 0.000 description 1
- 201000006938 muscular dystrophy Diseases 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 108010014614 prolyl-glycyl-proline Proteins 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000029983 protein stabilization Effects 0.000 description 1
- 230000020236 regulation of protein stability Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 238000010798 ubiquitination Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/95—Fusion polypeptide containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种增强目的蛋白降解的多肽。所述多肽信息包括编码多肽的cDNA序列，RNA序列和蛋白序列及其延伸序列、修饰过的氨基酸序列以及同5源性不低于70%的蛋白序列。本发明还提供了一种增强蛋白降解多肽的应用，通过与目的蛋白融合，该多肽可以特异增强自身降解并可以介导融合蛋白的共降解，其降解程度显著，可以达到近乎敲除的效果。该多肽可用做调控蛋白水平的工具，在体外或者体内调节融合蛋白的表达水平来研究其功能和调节生物学通路。该多肽可以通过体内和体外合成得到，方便快捷，为今后关于蛋白聚集疾病的治疗提供新的途径和方法。
Description
技术领域
本发明涉及基因工程技术领域，特别涉及一种增强蛋白降解的多肽及其应用。
背景技术
蛋白稳定性的调控发挥重要的生物学功能。在体内，蛋白稳定性失调与众多疾病相关。由于蛋白质聚集体的形成而促发的疾病被称为蛋白质聚集疾病。其中最常见的是神经退行性疾病，如蛋白质异常聚集并沉积在大脑的特定区域并阻碍其功能（参考文献：Protein aggregation and neurodegenerative diseases: From theory to therapy）。蛋白降解主要有蛋白酶体介导的泛素级联反应以及溶酶体介导的降解途径。其中泛素化在癌症，代谢综合症，神经退行性疾病，自身免疫疾病，炎症性失常，感染和肌肉营养不良的蛋白稳定调控方面发挥重要作用（参考文献：Ubiquitination in disease pathogenesisand treatment）。通过激活（E1），结合（E2）和连接（E3）酶进行的级联酶促反应，特异的蛋白序列被识别，标记，在特定信号通路的调解下，使得目的蛋白得到精确的降解（参考文献：The ubiquitin system）。特定的蛋白序列可以在体外作为一种标签，通过与要研究的目的蛋白融合介导其快速地酶解来研究其功能和调节生物学通路。直接靶向蛋白质的方法比靶向DNA和RNA的方法在在特异性，可逆性和时间方面上具有优势（参考文献：ConditionalDegrons for Controlling Protein Expression at the Protein Level）。
发明内容
本发明设计一种增强蛋白降解的多肽及其应用，具体为该多肽及同源多肽的序列，及作为标签，调节融合目的蛋白稳定性的方法。
本发明包括蛋白降解多肽J100的序列、包括J100的延伸序列、修饰过的氨基酸序列以及同源性不低于70%的蛋白序列。通过与目的蛋白融合，我们发现该降解多肽J100具有调控融合蛋白共同降解的功能，因此该降解多肽J100可以作为一种多肽标签，应用在分子生物学及细胞生物学领域。
附图说明
图1为J1-J6截短体的蛋白表达情况。图中，左侧为截短体western blot检测目的蛋白的表达；右侧为待检测蛋白的区域示意图，星号表示表达强度，灰色为表达，黑色为不表达。
具体实施方式
下文分别提供降解多肽J100鉴定过程及方法，以及与荧光蛋白tdTomato融合从而增强tdTomato降解的效果。降解多肽J100的鉴定过程及方法，以及对荧光蛋白的促降解作用，并不以任何方式限制本发明的范围。
1，降解多肽J100的鉴定及方法
HEK293细胞常规培养，培养条件为 5%二氧化碳培养箱，37度恒温。培养基为DMEM，双抗， 10%血清和L-谷氨酰胺。细胞生长到50%-80%时，进行转染，转染方法参照脂质体2000的说明，转染两到四天后收集细胞，裂解后按照每孔上样10 µg进行western blot，利用flag抗体进行检测目的蛋白的表达情况。
利用高通量筛选方法，我们发现一段蛋白序列J2 （图1 所示）显著影响蛋白稳定，Flag标签无法检测到J2的表达。通过截短体筛选，最终将影响蛋白稳定性的序列锁定到100aa内（见图1 中J5与J6的表达）。本实验鉴定到了降解多肽J100。
2，降解多肽J100对荧光蛋白tdTomato的促降解作用
实验方法与结果
HEK293细胞常规培养，培养条件为 5%二氧化碳培养箱，37度恒温。培养基为DMEM，双抗， 10%血清和L-谷氨酰胺。细胞生长到50%-80%时，进行转染，转染方法参照脂质体2000的说明。转染12-36小时后在荧光显微镜下进行观察tdTomato红色荧光，普通视野作为对照。
我们可以看到，在单独转染tdTomato的细胞内，能观测到红色荧光信号，在共表达降解多肽J100的细胞中，几乎没有红色荧光的表达。该数据证明降解多肽J100对tdTomato蛋白表达的促降解作用。
序列表
<110> 拜澳泰克生物医学科技（沈阳）有限公司
<120> 一种增强蛋白降解多肽的发现及应用
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 303
<212> DNA
<213> 小鼠(Mus musculus)
<400> 1
agaagactgg ccaacaagat cgagcacgag ctgagcagag gcagcttcca ccccgtgccc 60
accagaggca gcgccctgga gaccaccaag agccccctga tcatcgacaa gaacgagcac 120
ttcaccgtgt acagagaccc cgccctgatc ggcagcgaga ccggcgccaa ccacatcagc 180
cccttcctga gccagcaccc cttcagcctg cacagcagca gccacagaac ctgcctgaac 240
cccggcaccc accaccccgc cctgaccccc ggcccccacc tgctggccgg cagcaccagc 300
tga 303
<210> 2
<211> 303
<212> RNA
<213> 小鼠(Mus musculus)
<400> 2
agaagacugg ccaacaagau cgagcacgag cugagcagag gcagcuucca ccccgugccc 60
accagaggca gcgcccugga gaccaccaag agcccccuga ucaucgacaa gaacgagcac 120
uucaccgugu acagagaccc cgcccugauc ggcagcgaga ccggcgccaa ccacaucagc 180
cccuuccuga gccagcaccc cuucagccug cacagcagca gccacagaac cugccugaac 240
cccggcaccc accaccccgc ccugaccccc ggcccccacc ugcuggccgg cagcaccagc 300
uga 303
<210> 3
<211> 100
<212> PRT
<213> 小鼠(Mus musculus)
<400> 3
Arg Arg Leu Ala Asn Lys Ile Glu His Glu Leu Ser Arg Gly Ser Phe
1 5 10 15
His Pro Val Pro Thr Arg Gly Ser Ala Leu Glu Thr Thr Lys Ser Pro
20 25 30
Leu Ile Ile Asp Lys Asn Glu His Phe Thr Val Tyr Arg Asp Pro Ala
35 40 45
Leu Ile Gly Ser Glu Thr Gly Ala Asn His Ile Ser Pro Phe Leu Ser
50 55 60
Gln His Pro Phe Ser Leu His Ser Ser Ser His Arg Thr Cys Leu Asn
65 70 75 80
Pro Gly Thr His His Pro Ala Leu Thr Pro Gly Pro His Leu Leu Ala
85 90 95
Gly Ser Thr Ser
100
Claims (3)
1.一种增强蛋白降解的方法，其特征在于，多肽通过与目的蛋白融合并具有调控融合蛋白共同降解的功能，所述多肽为SEQ ID NO:3 所示的氨基酸序列。
2.根据权利要求1中所述的一种增强蛋白降解的方法，其特征在于，该多肽能与目的蛋白融合并调控共同降解。
3.根据权利要求1-2中任意一项所述的增强蛋白降解的方法，其特征在于，转染对象为真核细胞。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910383227.1A CN110092838B (zh) | 2019-05-09 | 2019-05-09 | 一种增强蛋白降解多肽的发现及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910383227.1A CN110092838B (zh) | 2019-05-09 | 2019-05-09 | 一种增强蛋白降解多肽的发现及应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110092838A CN110092838A (zh) | 2019-08-06 |
| CN110092838B true CN110092838B (zh) | 2023-03-10 |
Family
ID=67447409
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910383227.1A Active CN110092838B (zh) | 2019-05-09 | 2019-05-09 | 一种增强蛋白降解多肽的发现及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110092838B (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105177023A (zh) * | 2014-05-30 | 2015-12-23 | 华东理工大学 | 一种在真核细胞内其稳定性受蓝光调控的光敏降解子desVVD |
| WO2017079723A1 (en) * | 2015-11-07 | 2017-05-11 | Board Of Regents, The University Of Texas System | Targeting proteins for degradation |
| CN108239656A (zh) * | 2016-12-27 | 2018-07-03 | 天津天锐生物科技有限公司 | 一种小分子药物控制的蛋白功能开关系统 |
-
2019
- 2019-05-09 CN CN201910383227.1A patent/CN110092838B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105177023A (zh) * | 2014-05-30 | 2015-12-23 | 华东理工大学 | 一种在真核细胞内其稳定性受蓝光调控的光敏降解子desVVD |
| WO2017079723A1 (en) * | 2015-11-07 | 2017-05-11 | Board Of Regents, The University Of Texas System | Targeting proteins for degradation |
| CN108239656A (zh) * | 2016-12-27 | 2018-07-03 | 天津天锐生物科技有限公司 | 一种小分子药物控制的蛋白功能开关系统 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110092838A (zh) | 2019-08-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3487998B1 (en) | Compositions and methods for identifying rna binding polypeptide targets | |
| KR101481142B1 (ko) | 코리네박테리아 발현용 합성프로모터 | |
| US20190202856A1 (en) | Engineered crispr proteins for covalent tagging nucleic acids | |
| WO2021110119A1 (zh) | 一种高活性转座酶及其应用 | |
| Narayan et al. | Soluble expression, purification, and secondary structure determination of human PDX1 transcription factor | |
| Narayan et al. | Protein production and purification of a codon-optimized human NGN3 transcription factor from E. coli | |
| CN110092838B (zh) | 一种增强蛋白降解多肽的发现及应用 | |
| WO2024235228A1 (zh) | 一种性能提升的rna聚合酶突变体 | |
| EP4303310A1 (en) | Method for producing cas3 protein | |
| CN104568861A (zh) | 一种基于双分子互补的共价修饰标记来检测蛋白相互作用的方法 | |
| Xiong et al. | High efficiency and throughput system in directed evolution in vitro of reporter gene | |
| Finger et al. | Impact of rare codons and the functional coproduction of rate-limiting tRNAs on recombinant protein production in Bacillus megaterium | |
| CN114591987B (zh) | 一种用于检测活细胞中mTORC1活性的遗传编码荧光生物传感器及其构建方法 | |
| CN109880840A (zh) | 一种重组蛋白大肠杆菌体内生物素化标记系统 | |
| CN110093361B (zh) | 一种增强基因表达的增强子多肽及其应用 | |
| CN115261359A (zh) | 一种新型crispr酶和系统以及应用 | |
| CN114560950A (zh) | 一种遗传编码的有机汞荧光探针及其制备方法和应用 | |
| WO2021164665A1 (zh) | 一种精氨酸荧光探针及其制备方法和应用 | |
| KR102491095B1 (ko) | Cas 단백질 및 HIF의 TAD 도메인을 포함하는 단백질 융합체 및 이의 용도 | |
| US12054520B2 (en) | Control of and imaging techniques for protein nanoscaffolds | |
| KR100730875B1 (ko) | 번역반응 촉진 ｄｎａ 단편 및 그것을 사용한 무세포계단백질 합성방법 | |
| US12291729B2 (en) | Engineered Cas protein and use thereof | |
| CN115960924A (zh) | 重组jo-1抗原的制备方法和应用 | |
| Wheway | Genetic Engineering of Ciliated Cells | |
| Lotti et al. | Aggregation of recombinant proteins: Understanding basic issues to overcome production bottlenecks |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 110041 Dadong District, Shenyang City, Liaoning Province, 104 Northeast Da Avenue (Gate 1) Applicant after: Baiaotec (Shenyang) biomedical Group Co.,Ltd. Address before: 110041 Dadong District, Shenyang City, Liaoning Province, 104 Northeast Da Avenue (Gate 1) Applicant before: BIOTECH BIOMEDICAL TECHNOLOGY (SHENYANG) Co.,Ltd. |
|
| CB02 | Change of applicant information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |