Autophagic inhibiting polypeptide

A technology of autophagy inhibition and cell technology, applied in the field of bioengineering, can solve the problems of limiting the clinical application of chloroquine, poor prognosis of patients, low specificity of chloroquine inhibiting cell autophagy, etc., to achieve large-scale popularization and utilization, convenient operation, and beneficial Effects at the cellular level and at the animal level

Active Publication Date: 2017-12-22
江苏护理职业学院
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  • Abstract
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  • Claims
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Problems solved by technology

In addition, chloroquine has low specificity in inhibiting autophagy, and it also has other biological functions, such as chloroquine can prevent DNA replication and RNA

Method used

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  • Autophagic inhibiting polypeptide
  • Autophagic inhibiting polypeptide
  • Autophagic inhibiting polypeptide

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0021] Example 1

[0022] 1. The interaction test between FIP200 and its truncated mutants and the ATG13 protein, with the purpose of identifying the smallest structural domains that are potentially required to mediate FIP200 and ATG13 proteins:

[0023] Main experimental materials:

[0024] 1) Cell

[0025] In this experiment, 293T cells were selected. The cell culture is convenient, the growth is fast, and the liposome transfection in vitro has a high autophagy activity. In the case of autophagy induction, through immunoprecipitation, the interaction between FIP200 and ATG13 protein can be significantly detected. The label LC3ii band is easier to detect.

[0026] 293T, cells, purchased from ATCC, culture medium DMEM Gibico, containing 10% fetal bovine serum Life Tech, cultured in a cell incubator with 5% CO2, 37 degrees, 95% humidity, 3), antibody, flag antibody sigma, 1:2000, HA-beads sigma, GFP antibody, FIP200 antibody, LC3 antibody, cell signal technology company.

[0027] First...

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PUM

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Abstract

The invention relates to an autophagic inhibiting polypeptide which has an amino acid sequence of LPDISLKDLQFLQSFCPSEVQ. The polypeptide can be prepared by an artificial synthesis or host cell expression method, and can be used for preparing tumor treatment medicines. The autophagic inhibiting polypeptde only has 21 amino acids, can be easily and feasibly prepared by either artificially synthesizing or applying host cell expression, and is beneficial to large-scale popularization and utilization. The polypeptide is derived from inherent protein in a living body, cannot cause immune elimination in an animal experiment, has small toxicity to cells, and is more specific. The inhibiting polypeptide can be used for introducing a gene segment coding the polypeptide into a cell for stable expression through a lentivirus or transfection and other means, is convenient to operate, and is beneficial to researches for cell level and animal level.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the preparation and application of an autophagy "switch" initiation protein complex polypeptide inhibitor. Background technique [0002] The autophagy pathway is a new target for tumor drug therapy. The classic autophagy process can be seen in figure 1 shown. In recent years, the idea that cancer cells "eat themselves to survive" suggests that autophagy may play an important role in tumorigenesis and progression. For example, before the formation of abundant blood vessels around tumor cells, the nutrients produced by the autophagic degradation pathway can provide energy for tumor cells, thereby maintaining their survival under extreme stress and lack of nutrients. A large number of other studies have confirmed that the number of autophagosomes usually increases after the use of anticancer drugs and X-ray irradiation in some tumor cells, and the degree of increase is posi...

Claims

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Application Information

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IPC IPC(8): C07K14/47A61K38/17A61P35/00
CPCA61K38/00C07K14/4703
Inventor 陈松陈丹韦栋平季润元徐秉忠
Owner 江苏护理职业学院
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