4HVY

A thermostable variant of human NUDT18 NUDIX domain obtained by Hot Colony Filtration

Summary for 4HVY

• Entry DOI: 10.2210/pdb4hvy/pdb
• Related: 3GG6
• Descriptor: Nucleoside diphosphate-linked moiety X motif 18, GLYCEROL, MAGNESIUM ION, ... (6 entities in total)
• Functional Keywords: hydrolase, thermostability, directed evolution, hot-cofi, hot colony filtration
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 18761.17

Authors

Asial, I.,Cheng, Y.X.,Engman, H.,Wu, B.,Dollhopf, M.,Nordlund, P.,Cornvik, T. (deposition date: 2012-11-07, release date: 2014-01-01, Last modification date: 2024-02-28)

Primary citation

Asial, I.,Cheng, Y.X.,Engman, H.,Dollhopf, M.,Wu, B.,Nordlund, P.,Cornvik, T.
Engineering protein thermostability using a generic activity-independent biophysical screen inside the cell.
Nat Commun, 4:2901-2901, 2013

PubMed Abstract: Protein stability is often a limiting factor in the development of commercial proteins and biopharmaceuticals, as well as for biochemical and structural studies. Unfortunately, identifying stabilizing mutations is not trivial since most are neutral or deleterious. Here we describe a high-throughput colony-based stability screen, which is a direct and biophysical read-out of intrinsic protein stability in contrast to traditional indirect activity-based methods. By combining the method with a random mutagenesis procedure, we successfully identify thermostable variants from 10 diverse and challenging proteins, including several biotechnologically important proteins such as a single-chain antibody, a commercial enzyme and an FDA-approved protein drug. We also show that thermostabilization of a protein drug using our approach translates into dramatic improvements in long-term stability. As the method is generic and activity independent, it can easily be applied to a wide range of proteins.

PubMed: 24352381
DOI: 10.1038/ncomms3901

Experimental method

X-RAY DIFFRACTION (1.46 Å)