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Crystal of rat RANK/RANKL extracellular compound, preparation method and application thereof

A complex and crystal technology, which is applied in the field of purifying and expressing RANK extracellular protein and preparing mouse RANK/RANKL extracellular complex crystals, which can solve the problems of difficult to obtain crystals and large variability.

Inactive Publication Date: 2010-09-29
GENERAL HOSPITAL OF PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

2. Compared with the relatively rigid structure of TNF protein family members including RANKL, the structure of TNFR protein family members including RANK has greater variability, so it is more difficult to obtain crystallization

Method used

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  • Crystal of rat RANK/RANKL extracellular compound, preparation method and application thereof
  • Crystal of rat RANK/RANKL extracellular compound, preparation method and application thereof
  • Crystal of rat RANK/RANKL extracellular compound, preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0031] 1. Expression and purification

[0032] 1. Expression and purification of mouse RANK extracellular domain protein

[0033] The amino acid sequence of mouse RANK is as follows:

[0034] MAPRARRRRQLPAPLLALCVLLVPL QVTLQVTPPCTQERHYEHLGRCCSR CEPGKYLSSKCTPTSDSVCLPCGPDEYLDTWNEEDKCLLHKVCDAGKALV AVDPGNHTAPRRCACTAGYHWNSDCECCRRNTECAPGFGAQHPLQLNKDT VCTPCLLGFFSDVFSSTDKCKPWTNCTLLGKLEAHQGTTESDVVCSSSMT LRRPPKEAQA YLPSLIVLLLFISVVVVAAIIFGVYYRKGGKALTANLWNWVNDACSSLSGNKESSGDRCAGSHSATSSQQEVCEGILLMTREEKMVPEDGAGVCGPVCAAGGPWAEVRDSRTFTLVSEVETQGDLSRKIPTEDEYTDRPSQPSTGSLLLIQQGSKSIPPFQEPLEVGENDSLSQCFTGTESTVDSEGCDFTEPPSRTDSMPVSPEKHLTKEIEGDSCLPWVVSSNSTDGYTGSGNTPGEDHEPFPGSLKCGPLPQCAYSMGFPSEAAASMAEAGVRPQDRADERGASGSGSSPSDQPPASGNVTGNSNSTFISSGQVMNFKGDIIVVYVSQTSQEGPGSAEPESEPVGRPVQEETLAHRDSFAGTAPRFPDVCATGAGLQEQGAPRQKDGTSRPVQEQGGAQTSLHTQGSGQCAE

[0035] The underlined part is the amino acid sequence of the extracellular region of mouse RANK

[0036] Material

[0037] Oligonucleotides were prepared...

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Abstract

The invention provides a method for purifying and expressing RANK extracellular area protein, and also provides a method for preparing RANK / RANKL compound crystals, a three-dimensional crystal structure of the RANK / RANKL compound, and applications of the crystal in the medical field.

Description

technical field [0001] The invention relates to a method for purifying and expressing RANK extracellular protein, and also provides a crystal of RANK / RANKL complex, especially a crystal formed of mouse RANK / RANKL extracellular protein. The invention provides a method for preparing rat RANK / RANKL extracellular complex crystal and the application of the complex structure in the field of medicine. Background technique [0002] Cytokines in the tumor necrosis factor (TNF) family play important roles in various biological functions such as inflammatory response, organogenesis, host defense, autoimmunity and apoptosis. These biological regulatory factors play corresponding physiological roles by triggering intracellular signal transmission and changing cell phenotype through receptor-ligand interaction. As a member of the TNF family, RANKL and its receptor RANK play an important role in various activities in the body, and play an important role in regulating the bone remodeling p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/70C12N15/74C12P21/02C30B29/54G01N33/68C12R1/19
Inventor 唐佩福刘长振黄鹏张世谦高斌
Owner GENERAL HOSPITAL OF PLA
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