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Modified adiponectin polypeptides and their uses

A technology of adiponectin and modifiers, applied in the field of adiponectin polypeptides, can solve problems such as lack of reproducibility, loss of protein activity, and slow reaction of derivatives

Inactive Publication Date: 2015-01-21
AMBRX
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some derivatives react too slowly to be practically usable
Some derivatives cause loss of protein activity due to attachment to sites responsible for protein activity
Some derivatives are not specific for the site to which they will be attached, which also leads to loss of desired activity and lack of reproducibility of results

Method used

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  • Modified adiponectin polypeptides and their uses
  • Modified adiponectin polypeptides and their uses
  • Modified adiponectin polypeptides and their uses

Examples

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example

[0634] The following examples are offered to illustrate, not limit, the claimed invention.

example 1

[0636] This example describes one of several potential sets of criteria for selecting preferred sites for incorporation of non-naturally encoded amino acids into adiponectin.

[0637] This example illustrates how to select preferred sites within the adiponectin polypeptide for introducing non-naturally encoded amino acids. The crystal structures of adiponectin, adiponectin dimers, adiponectin trimers, and adiponectin multimers were used to determine preferred locations at which one or more non-naturally encoded amino acids may be introduced. Other adiponectin structures (eg, from computer modeling, etc.) can be employed to examine potential variations in major and minor structural elements between crystal structure datasets. The coordinates of these structures can be purchased from the Protein Data Bank (PDB) (Berstein (Berstein) et al. J. Molecular Biology 1997, 112, p. 535) or through the information available on the World Wide Web at rcsb.org. Acquired from The Research Co...

example 2

[0643] This example details the cloning and expression of hGH polypeptides including non-naturally encoded amino acids in E. coli. This example also describes a method for assessing the biological activity of modified hGH polypeptides, and these cloning, expression and assessment techniques can also be used for adiponectin polypeptides of the invention.

[0644] Methods for cloning hGH and fragments thereof are described in detail in US Patent Nos. 4,601,980; 4,604,359; 4,634,677; 4,658,021; 4,898,830; The sequences of hGH or the mature form of hGH lacking the N-terminal signal sequence are included in Patent Nos. 4,601,980; 4,604,359; 4,634,677; 4,658,021; 4,898,830; , each incorporated herein by reference for all purposes.

[0645] hGH containing non-naturally encoded amino acids was expressed using an introduced translation system comprising an orthogonal tRNA (O-tRNA) and an orthogonal aminoacyl tRNA synthetase (O-RS). O-RS preferentially aminoacylates O-tRNAs with non-n...

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Abstract

Modified adiopnectin polypeptides and uses thereof are provided.

Description

technical field [0001] The present invention relates to adiponectin polypeptides modified with at least one non-naturally encoded amino acid. Background technique [0002] Adiponectin is a protein that regulates several metabolic processes including glucose regulation and fatty acid breakdown. Adiponectin is secreted into the bloodstream by adipose tissue and in adults, plasma levels of the hormone are inversely correlated with body fat percentage. The hormone acts to suppress metabolic disturbances that lead to type 2 diabetes, obesity, atherosclerosis and non-alcoholic fatty liver disease (NAFLD). [0003] Adiponectin is also known as AdipoQ, apM1, GBP28 or Acrp30 (Scherer et al., J Biol Chem, 270:26746-9, 1995; Hu et al., J Biol Chem., 271:10697-703, 1996; Maeda et al., Biochem Biophys Res Commun, 221:286-9, 1996; and Nakano et al., Biochemical Journal (Tokyo ), 120:803-12, 1996), is a polypeptide composed of an N-terminal collagen domain and a C-terminal globular doma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00
CPCC07K14/5759A61K38/00C07K14/575
Inventor 瓦迪姆·克赖诺夫安娜-玛丽亚·A.·派特南
Owner AMBRX