Riken Corporation

Japan Science and Technology Agency

National Food Research Institute

Other

Sysmex Corporation

Hokkaido University

Mochida Pharmaceutical Co., Ltd.

The Jikei University

Riken Japan Science and Technology Agency of 1-8

Mikako Shirouzu

Shigeyuki Yokoyama

Takanori Kigawa

Namtip Chumpolkulwong

Kensaku Sakamoto

Kozo Ochi

Chie Hori

Goushi Ishihara

Kaori Tajima

Ayako Sakamoto

Mie Goto

Takeshi Hosaka

Toshitada Takemori

Akiko Tanaka

Takashi Saito

Osamu Nureki

Yoshihiro Nakamura

Kazuki Saito

Ryuichiro Ishitani

Takashi Yabuki

Soichi Kojima

Tomokazu Matsuura

Natsuko Matsuda

Takehisa Matsumoto

Tetsuya Hori

Hiroaki Tanabe

Miho Tanaka

Naoko Nomura

Kaori Ito

Yoshiko Katsura

Takehiro Shinoda

Tomomi Someya

Mihoro Saeki

Hideo Ogiso

Chie Takemoto

Kozo Suto

Kazumi Shimono

Tamami Ueshima

Toru Kondo

Ikuyo Inoue

Chiemi Tsumagari

Takahiro Masaki

Masahiro Miura

In order to provide a membrane protein production method which does not require the step of solubilizing a membrane protein and which allows the membrane protein having an excellent quality to be obtained with a high yield, a method in accordance with an embodiment of the present invention includes: a step (a) of preparing a reaction solution for cell-free protein synthesis, the reaction solution containing (i) a template nucleic acid which encodes the membrane protein, (ii) a lipid, and (iii) a detergent which is contained at a concentration equal to or higher than a critical micelle concentration; and a step (b) of synthesizing the membrane protein while the concentration of the detergent in the reaction solution is maintained at a concentration equal to or higher than a critical micelle concentration.

Method of manufacturing membrane protein and utilization thereof

In order to provide an antibody having high therapeutic and prophylactic effects against cancer and the like, three types of mouse monoclonal antibodies were prepared which exhibit high affinities for a human-derived Eva1 protein. Moreover, constant regions of these antibodies were substituted with human-derived constant regions to also prepare chimeric antibodies. Further, these mouse antibodies and chimeric antibodies were found to have high ADCC and/or CDC activities. Furthermore, it was also revealed that administering these antibodies to mice having been subjected to melanoma cell administration suppresses the metastasis and the like of the cells to the lungs.

Anti-Eva1 protein antibody

Disclosed is an isolated monoclonal antibody against a human Transforming Growth Factor-&#x3b2; (TGF-&#x3b2;) Latency Associated Protein (LAP) degradate, the isolated monoclonal antibody being capable of recognizing an integrin binding site in the human TGF-&#x3b2; LAP degradate.

Antibody against human TGF-&#x3b2; LAP degradate, and use thereof

It is intended to provide an extract, kit and a process for synthesizing a protein in a cell-free system. The extract is characterized by being prepared from mutantcells having a mutation in the ribosomal protein S12 gene. In a preferred embodiment, the mutation is such a mutation as conferring a resistance or dependence to streptomycin onwhereby the reading efficiency of mRNA codon on ribosomes can be improved and thus the protein productivity can be elevated.

[object Object]

A method is provided for producing a membrane protein folded to its native structure or active structure in a lipid disc or a liposome. The method comprises: (a) preparing a reaction solution for cell-free protein synthesis containing a polynucleotide encoding a membrane protein, a steroidal detergent, and a phospholipid, wherein the steroidal detergent is contained at a concentration higher than its critical micelle concentration; (b) decreasing the concentration of said steroidal detergent in the reaction solution; and (c) synthesizing the membrane protein simultaneously with formation of a lipid disk or liposome into which the synthesized membrane protein is integrated.

Method of using cell-free protein synthesis to produce a membrane protein

The present invention discloses an expression method for non-naturally-occurring amino acid-containing protein comprising: expressing in animal cells: (A) a mutant tyrosyl-tRNA synthetase that is a mutation of tyrosyl-tRNA synthetase originating inwith an enhanced specificity for a non-naturally-occurring tyrosine derivative as compared with the specificity for tyrosine; (B) suppressor tRNA originating inspecies,species orspecies of eubacteria and capable of binding with the tyrosine derivative in the presence of the mutant tyrosyl tRNA synthetase; and, (C) a desired protein gene that has undergone a nonsense mutation at a desired site; wherein, the tyrosine derivative is incorporated at the location of this nonsense mutation.

DNA sequences for the expression of alloproteins

It is intended to provide a process for producing an extract for cell-free protein synthesis whereby the productivity of a protein and the production efficiency can be improved. Cells are cultured under suppressed growth conditions. In the stationary phase of the culture, the cells are collected and then disrupted. The above-described cells are preferably bacterial cells, in particular,cells. In the suppressed growth conditions as described above, the culture temperature is preferably from 20 to 32&#xb0; C., more preferably 26&#xb0; C. or higher and lower than 30&#xb0; C.

Process for producing extract for cell-free protein synthesis and cell extract produced thereby

A method of expressing a protein having an unnatural amino acid integrated thereinto comprising expressing (A) a mutant tyrosyl tRNA synthase that is derived from-origin tyrosyl tRNA synthase and has an elevated specificity for an unnatural tyrosine derivative compared with the specificity for tyrosine, (B) a suppressor tRNA originating in an eubacterium belonging to the genusorwhich is capable of binding to the above tyrosine derivative in the presence of the above mutant TyrRS, and (C) a desired protein gene having a nonsense mutation at a desired site in animal cells to thereby incorporate the above tyrosine derivative into the nonsense mutation site in the above protein.

Method of expressing proteins comprising non-naturally-occurring amino acids

The present invention relates to a crystal of a complex of an epidermal growth factor (EGF) and an epidermal growth factor receptor (EGFR), a crystal of a complex of EGFR and a substance regulating EGFR activity, structure coordinates of these crystals, a method for screening for the substance regulating EGFR activity, a method for designing the substance regulating EGFR activity, a method for designing an EGFR variant or an EGF variant, a method for producing an EGFR variant or an EGF variant and an EGF variant or an EGFR variant obtainable by such method, a method for designing an epitope using the structure coordinates of the EGF-EGFR complex, a method for producing an anti-EGFR antibody or an anti-EGF antibody and an antibody obtainable by such method, a polypeptide or a salt thereof comprising a region that forms an EGFR dimerization site, and the like.

EGR-EGFR complex

We claim a method of expression a thioredoxin-fused membrane protein comprising expressing said thioredoxin-fused membrane protein in a cell-free protein synthesis system in the presence of a non-ionic detergent. It is preferable that the fused membrane protein which is a highly hydrophobic protein, e.g., a G protein-coupled receptor, can be produced in a state being highly soluble and capable of forming a biologically active three-dimensional structure. Also, this invention provides a recombinant polynucleotide encoding said thioredoxin-fused membrane protein which is expressed in the cell-free protein synthesis system.

Method for producing by cell-free protein synthesis system using thioredoxin-fused protein expression vector

A method for producing a protein using a cell-free protein synthesis system comprising a detergent so that the protein can be synthesized without aggregation, is provided. The protein is a protein comprising a hydrophobic region in at least a portion thereof, for example, a membrane protein or its fragment (portion). And the detergent is a mild detergent which would not denature the protein, for example, a nonionic or amphoteric ionic detergent.

Process for synthesizing protein using cell-free protein synthesis system

Growing community of inventors

Yokohama, Japan

Mikako Shirouzu