The conformation and stability of recombinant-derived granulocyte-macrophage colony stimulating factors.


Abstract

The conformation and stability of recombinant-derived human and murine granulocyte-macrophage colony stimulating factors produced in Escherichia coli have been investigated by analytical ultracentrifugation, urea-gradient polyacrylamide gel electrophoresis and several spectroscopic methods. The proteins were demonstrated to be physically homogeneous monomeric proteins with compact globular shapes and shown to have similar secondary structures containing both alpha-helix and beta-sheet structure. The intramolecular disulphide linkages of both proteins were shown to be essential for maintaining native conformation as reduction with dithiothreitol resulted in protein unfolding. Comparison of the human E. coli-derived (non-glycosylated) and mammalian cell culture-derived (glycosylated) proteins by urea-gradient electrophoresis indicated that glycosylation had no major effect on the conformational stability and kinetics of urea induced unfolding and refolding. Study holds ProTherm entries: 7550, 7551, 7552, 7553 Extra Details:

Submission Details

ID: zXr5fXfp3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:34 p.m.

Version: 1

Publication Details
Wingfield P;Graber P;Moonen P;Craig S;Pain RH,Eur. J. Biochem. (1988) The conformation and stability of recombinant-derived granulocyte-macrophage colony stimulating factors. PMID:3281835
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
100.0 Granulocyte-macrophage colony-stimulating factor P01587 CSF2_MOUSE
94.4 Granulocyte-macrophage colony-stimulating factor Q0MUT8 CSF2_CHLAE
100.0 Granulocyte-macrophage colony-stimulating factor P04141 CSF2_HUMAN