Reengineering granulocyte colony-stimulating factor for enhanced stability.


Abstract

Granulocyte colony-stimulating factor is a long-chain cytokine that has both biological and therapeutic applications. It is involved in the production and maturation of neutrophilic progenitor cells and neutrophils and is administered to stimulate the production of white blood cells to reduce the risk of serious infection in immunocompromised patients. We have reengineered granulocyte colony-stimulating factor to improve the thermodynamic stability of the protein, focusing on enhancing the alpha-helical propensity of residues in the antiparallel 4-helix bundle of the protein. These redesigns resulted in proteins with substantially enhanced stability while retaining wild-type levels of biological activity, measured as the ability of the reengineered proteins to stimulate the proliferation of murine myeloid cells transfected with the granulocyte colony-stimulating factor receptor. Study holds ProTherm entries: 11566, 11567, 11568, 11569, 11570, 11571, 11572, 11573, 11574, 11575, 11576, 11577 Extra Details: cytokine; alpha-helical propensity; 4-helix bundle;,biological activity

Submission Details

ID: gjBTbEyP3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:42 p.m.

Version: 1

Publication Details
Bishop B;Koay DC;Sartorelli AC;Regan L,J. Biol. Chem. (2001) Reengineering granulocyte colony-stimulating factor for enhanced stability. PMID:11406632
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1GNC 1994-07-31 STRUCTURE AND DYNAMICS OF THE HUMAN GRANULOCYTE COLONY-STIMULATING FACTOR DETERMINED BY NMR SPECTROSCOPY. LOOP MOBILITY IN A FOUR-HELIX-BUNDLE PROTEIN
5GW9 2017-09-13 1.65 Crystal structure of C163, a backbone circularized G-CSF
5ZO6 2019-04-17 1.7 Crystal structure of C166, a backbone circularized G-CSF
6BFS 2018-09-12 2.0 The mechanism of GM-CSF inhibition by human GM-CSF auto-antibodies
5D70 2015-10-14 2.06 Crystal structure of MOR03929, a neutralizing anti-human GM-CSF antibody Fab fragment in complex with human GM-CSF
1RHG 1994-01-31 2.2 THE STRUCTURE OF GRANULOCYTE-COLONY-STIMULATING FACTOR AND ITS RELATIONSHIP TO THOSE OF OTHER GROWTH FACTORS
5D71 2015-10-14 2.25 Crystal structure of MOR04302, a neutralizing anti-human GM-CSF antibody Fab fragment in complex with human GM-CSF
2GMF 1996-11-08 2.4 HUMAN GRANULOCYTE MACROPHAGE COLONY STIMULATING FACTOR
5D72 2015-10-14 2.6 Crystal structure of MOR04252, a neutralizing anti-human GM-CSF antibody Fab fragment in complex with human GM-CSF
6BFQ 2018-09-12 2.6 The mechanism of GM-CSF inhibition by human GM-CSF auto-antibodies
4RS1 2015-11-11 2.68 Crystal structure of receptor-cytokine complex
1CSG 1993-10-31 2.7 Three-dimensional structure of recombinant human granulocyte-macrophage colony-stimulating factor
2D9Q 2006-02-07 2.8 Crystal Structure of the Human GCSF-Receptor Signaling Complex
1CD9 2000-03-08 2.8 2:2 COMPLEX OF G-CSF WITH ITS RECEPTOR
5C7X 2015-10-14 2.95 Crystal structure of MOR04357, a neutralizing anti-human GM-CSF antibody Fab fragment in complex with human GM-CSF
4NKQ 2015-09-23 3.3 Structure of a Cytokine Receptor Complex
1PGR 2000-03-08 3.5 2:2 COMPLEX OF G-CSF WITH ITS RECEPTOR

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
98.3 Granulocyte colony-stimulating factor P09919 CSF3_HUMAN
94.4 Granulocyte-macrophage colony-stimulating factor Q0MUT8 CSF2_CHLAE
100.0 Granulocyte-macrophage colony-stimulating factor P04141 CSF2_HUMAN