The N-terminal domain of a tick evasin is critical for chemokine binding and neutralization and confers specific binding activity to other evasins

The Journal of Biological Chemistry
James R O EatonShoumo Bhattacharya

Abstract

Tick chemokine-binding proteins (evasins) are an emerging class of biologicals that target multiple chemokines and show anti-inflammatory activities in preclinical disease models. Using yeast surface display, we identified a CCL8-binding evasin, P672, from the tick Rhipicephalus pulchellus We found that P672 binds CCL8 and eight other CC-class chemokines with a Kd < 10 nm and four other CC chemokines with a Kd between 10 and 100 nm and neutralizes CCL3, CCL3L1, and CCL8 with an IC50 < 10 nm The CC chemokine-binding profile was distinct from that of evasin 1 (EVA1), which does not bind CCL8. We also show that P672's binding activity can be markedly modulated by the location of a StrepII-His purification tag. Combining native MS and bottom-up proteomics, we further demonstrated that P672 is glycosylated and forms a 1:1 complex with CCL8, disrupting CCL8 homodimerization. Homology modeling of P672 using the crystal structure of the EVA1 and CCL3 complex as template suggested that 44 N-terminal residues of P672 form most of the contacts with CCL8. Replacing the 29 N-terminal residues of EVA1 with the 44 N-terminal residues of P672 enabled this hybrid evasin to bind and neutralize CCL8, indicating that the CCL8-binding properties of...Continue Reading

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Citations

Jun 7, 2019·The Journal of Biological Chemistry·Angela W LeeShoumo Bhattacharya
May 29, 2020·Proceedings of the National Academy of Sciences of the United States of America·Charlotte FranckRichard J Payne
Jul 25, 2019·Frontiers in Physiology·Iveta ŠtibrániováMária Kazimírová
Nov 16, 2018·Parasites & Vectors·Pavlina VechtovaLibor Grubhoffer
Jun 27, 2019·The Journal of Biological Chemistry·Stepan S DenisovIngrid Dijkgraaf
May 31, 2020·The Journal of Biological Chemistry·Benoit DarlotShoumo Bhattacharya
Feb 13, 2020·PLoS Neglected Tropical Diseases·Tae Kwon KimAlbert Mulenga
Aug 21, 2020·The Journal of Biological Chemistry·Stepan S DenisovIngrid Dijkgraaf
Feb 23, 2020·Journal of Proteome Research·Shayli Varasteh MoradiKirill Alexandrov
Nov 9, 2021·Frontiers in Cellular and Infection Microbiology·Shoumo Bhattacharya, Patricia Anne Nuttall

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Datasets Mentioned

BETA
JAA60789.1
EZ406190.1
PXD008025

Methods Mentioned

BETA
fluorescence-activated cell sorting
FACS
surface display
glycosylation
size exclusion chromatography
biolayer interferometry
biolayer
surface
PCR
electrophoresis

Software Mentioned

Arpeggio
FigTree
PyMod
SignalP
MODELLER
Arpeggio server
ExPASy
EMBOSS
Xcalibur
Plotcon

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