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@ARTICLE{Ciepluch:860022,
author = {Ciepluch, Karol and Radulescu, Aurel and Hoffmann, Ingo and
Raba, Andreas and Allgaier, Jürgen and Richter, Dieter and
Biehl, Ralf},
title = {{I}nfluence of {PEG}ylation on {D}omain {D}ynamics of
{P}hosphoglycerate {K}inase: {PEG} {A}cts {L}ike {E}ntropic
{S}pring for the {P}rotein},
journal = {Bioconjugate chemistry},
volume = {29},
number = {6},
issn = {1520-4812},
address = {Columbus, Ohio},
publisher = {American Chemical Society},
reportid = {FZJ-2019-00819},
pages = {1950 - 1960},
year = {2018},
abstract = {Protein–polymer conjugation is a widely used technique to
develop protein therapeutics with improved pharmacokinetic
properties as prolonged half-life, higher stability, water
solubility, lower immunogenicity, and antigenicity.
Combining biochemical methods, small angle scattering
(SAXS/SANS), and neutron spin–echo spectroscopy, here we
examine the impact of PEGylation (i.e., the covalent
conjugation with poly(ethylene glycol) or PEG) on structure
and internal domain dynamics of phosphoglycerate kinase
(PGK) to elucidate the reason for reduced activity that is
connected to PEGylation. PGK is a protein with a hinge
motion between the two main domains that is directly related
to function. We find that secondary structure and ligand
access to the binding sites are not affected. The ligand
induced cleft closing is unchanged. We observe an additional
internal motion between covalent bonded PEG and the protein
compatible with Brownian motion of PGK in a harmonic
potential. Entropic interaction with the full PEG chain
leads to a force constant of about 8 pN/nm independent of
PEG chain length. This additional force preserves protein
structure and has negligible effects on the functional
domain dynamics of the protein. PEGylation seems to reduce
activity just by acting as a local crowder for the ligands.
The newly identified interaction mechanism might open
possibilities to improve rational design of
protein–polymer conjugates.},
cin = {Neutronenstreuung ; JCNS-1 / ICS-1 / JCNS-FRM-II},
ddc = {540},
cid = {I:(DE-Juel1)JCNS-1-20110106 / I:(DE-Juel1)ICS-1-20110106 /
I:(DE-Juel1)JCNS-FRM-II-20110218},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551) /
6G4 - Jülich Centre for Neutron Research (JCNS) (POF3-623)
/ 6215 - Soft Matter, Health and Life Sciences (POF3-621)},
pid = {G:(DE-HGF)POF3-551 / G:(DE-HGF)POF3-6G4 /
G:(DE-HGF)POF3-6215},
experiment = {EXP:(DE-MLZ)KWS2-20140101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:29847099},
UT = {WOS:000436211700019},
doi = {10.1021/acs.bioconjchem.8b00203},
url = {https://juser.fz-juelich.de/record/860022},
}