Native electrospray mass spectrometry approaches to probe the interaction between zinc and an anti-angiogenic peptide from histidine-rich glycoprotein

Esther M. Martin; Frances D. L.  Kondrat; Alan J. Stewart; James H. Scrivens; Peter J. Sadler; Claudia A. Blindauer

doi:10.1038/s41598-018-26924-1

Native electrospray mass spectrometry approaches to probe the interaction between zinc and an anti-angiogenic peptide from histidine-rich glycoprotein

Esther M. Martin, Frances D. L. Kondrat, Alan J. Stewart, James H. Scrivens, Peter J. Sadler, Claudia A. Blindauer

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Abstract

Zinc modulates the biological function of histidine-rich glycoprotein (HRG) through binding to its His-rich region (HRR). The Zn²⁺-binding properties of a 35 amino-acid biologically-active peptide mimic of the HRR, HRGP330, were investigated using dissociative mass spectrometry approaches in addition to travelling-wave ion mobility mass spectrometry (TWIM-MS). Native mass spectrometry confirmed zinc binding to HRGP330; however, broadening of the ¹H NMR resonances upon addition of Zn²⁺ ions precluded the attainment of structural information. A complementary approach employing TWIM-MS indicated that HRGP330 has a more compact structure in the presence of Zn²⁺ ions. Top-down MS/MS data supported a metal-binding-induced conformational change, as fewer fragments were observed for Zn²⁺-bound HRGP330. Zn²⁺-bound fragments of both N-terminal and C-terminal ends of the peptide were identified from collision-induced dissociation (CID) and electron transfer dissociation/proton transfer reaction (ETD/PTR) experiments, suggesting that multiple binding sites exist within this region of HRG. The combination of mass spectrometry and NMR approaches provides new insight into the highly dynamic interaction between zinc and this His-rich peptide.

Original language	English
Article number	8646
Number of pages	13
Journal	Scientific Reports
Volume	8
DOIs	https://doi.org/10.1038/s41598-018-26924-1
Publication status	Published - 5 Jun 2018

Keywords

Ion mobility mass spectrometry
Top-down mass spectrometry
Zinc
Histidine-rich glycoprotein

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Martin-2018-Native-electrospray-SciRep-8646
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Zinc-dependant heparin neutralisation: Charecterisation of zinc-dependant heparin neutralisation by fibrinogen and histidine-rich glycoprotein.
Stewart, A. J. (PI) & Pitt, S. J. (CoI)
27/10/15 → 26/10/18
Project: Studentship
Role of zinc in controlling histidine: Role of zinc in controlling histidine-rich glycoprotein complex formation: Implications for the development of thrombotic complications.
Stewart, A. J. (PI), Naismith, J. (CoI) & Pitt, S. J. (CoI)
8/06/15 → 7/06/18
Project: Standard
Dynamics of Circulatory Zinc Handling: The Dynamics of Circulatory Zinc Handling and Transport
Stewart, A. J. (PI)
BBSRC
9/04/12 → 8/04/15
Project: Standard

Cite this

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title = "Native electrospray mass spectrometry approaches to probe the interaction between zinc and an anti-angiogenic peptide from histidine-rich glycoprotein",

abstract = "Zinc modulates the biological function of histidine-rich glycoprotein (HRG) through binding to its His-rich region (HRR). The Zn2+-binding properties of a 35 amino-acid biologically-active peptide mimic of the HRR, HRGP330, were investigated using dissociative mass spectrometry approaches in addition to travelling-wave ion mobility mass spectrometry (TWIM-MS). Native mass spectrometry confirmed zinc binding to HRGP330; however, broadening of the 1H NMR resonances upon addition of Zn2+ ions precluded the attainment of structural information. A complementary approach employing TWIM-MS indicated that HRGP330 has a more compact structure in the presence of Zn2+ ions. Top-down MS/MS data supported a metal-binding-induced conformational change, as fewer fragments were observed for Zn2+-bound HRGP330. Zn2+-bound fragments of both N-terminal and C-terminal ends of the peptide were identified from collision-induced dissociation (CID) and electron transfer dissociation/proton transfer reaction (ETD/PTR) experiments, suggesting that multiple binding sites exist within this region of HRG. The combination of mass spectrometry and NMR approaches provides new insight into the highly dynamic interaction between zinc and this His-rich peptide. ",

keywords = "Ion mobility mass spectrometry, Top-down mass spectrometry, Zinc, Histidine-rich glycoprotein",

author = "Martin, {Esther M.} and Kondrat, {Frances D. L.} and Stewart, {Alan J.} and Scrivens, {James H.} and Sadler, {Peter J.} and Blindauer, {Claudia A.}",

note = "This work was supported by the BBSRC (grant ref. BB/J006467/1 and CASE studentship to E.M.M.) and the British Heart Foundation (grant ref. PG/15/9/31270 and FS/15/42/31556).",

year = "2018",

month = jun,

day = "5",

doi = "10.1038/s41598-018-26924-1",

language = "English",

volume = "8",

journal = "Scientific Reports",

issn = "2045-2322",

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T1 - Native electrospray mass spectrometry approaches to probe the interaction between zinc and an anti-angiogenic peptide from histidine-rich glycoprotein

AU - Martin, Esther M.

AU - Kondrat, Frances D. L.

AU - Stewart, Alan J.

AU - Scrivens, James H.

AU - Sadler, Peter J.

AU - Blindauer, Claudia A.

N1 - This work was supported by the BBSRC (grant ref. BB/J006467/1 and CASE studentship to E.M.M.) and the British Heart Foundation (grant ref. PG/15/9/31270 and FS/15/42/31556).

PY - 2018/6/5

Y1 - 2018/6/5

N2 - Zinc modulates the biological function of histidine-rich glycoprotein (HRG) through binding to its His-rich region (HRR). The Zn2+-binding properties of a 35 amino-acid biologically-active peptide mimic of the HRR, HRGP330, were investigated using dissociative mass spectrometry approaches in addition to travelling-wave ion mobility mass spectrometry (TWIM-MS). Native mass spectrometry confirmed zinc binding to HRGP330; however, broadening of the 1H NMR resonances upon addition of Zn2+ ions precluded the attainment of structural information. A complementary approach employing TWIM-MS indicated that HRGP330 has a more compact structure in the presence of Zn2+ ions. Top-down MS/MS data supported a metal-binding-induced conformational change, as fewer fragments were observed for Zn2+-bound HRGP330. Zn2+-bound fragments of both N-terminal and C-terminal ends of the peptide were identified from collision-induced dissociation (CID) and electron transfer dissociation/proton transfer reaction (ETD/PTR) experiments, suggesting that multiple binding sites exist within this region of HRG. The combination of mass spectrometry and NMR approaches provides new insight into the highly dynamic interaction between zinc and this His-rich peptide.

AB - Zinc modulates the biological function of histidine-rich glycoprotein (HRG) through binding to its His-rich region (HRR). The Zn2+-binding properties of a 35 amino-acid biologically-active peptide mimic of the HRR, HRGP330, were investigated using dissociative mass spectrometry approaches in addition to travelling-wave ion mobility mass spectrometry (TWIM-MS). Native mass spectrometry confirmed zinc binding to HRGP330; however, broadening of the 1H NMR resonances upon addition of Zn2+ ions precluded the attainment of structural information. A complementary approach employing TWIM-MS indicated that HRGP330 has a more compact structure in the presence of Zn2+ ions. Top-down MS/MS data supported a metal-binding-induced conformational change, as fewer fragments were observed for Zn2+-bound HRGP330. Zn2+-bound fragments of both N-terminal and C-terminal ends of the peptide were identified from collision-induced dissociation (CID) and electron transfer dissociation/proton transfer reaction (ETD/PTR) experiments, suggesting that multiple binding sites exist within this region of HRG. The combination of mass spectrometry and NMR approaches provides new insight into the highly dynamic interaction between zinc and this His-rich peptide.

KW - Ion mobility mass spectrometry

KW - Top-down mass spectrometry

KW - Zinc

KW - Histidine-rich glycoprotein

U2 - 10.1038/s41598-018-26924-1

DO - 10.1038/s41598-018-26924-1

M3 - Article

SN - 2045-2322

VL - 8

JO - Scientific Reports

JF - Scientific Reports

M1 - 8646

ER -

Native electrospray mass spectrometry approaches to probe the interaction between zinc and an anti-angiogenic peptide from histidine-rich glycoprotein

Abstract

Keywords

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Projects

Zinc-dependant heparin neutralisation: Charecterisation of zinc-dependant heparin neutralisation by fibrinogen and histidine-rich glycoprotein.

Role of zinc in controlling histidine: Role of zinc in controlling histidine-rich glycoprotein complex formation: Implications for the development of thrombotic complications.

Dynamics of Circulatory Zinc Handling: The Dynamics of Circulatory Zinc Handling and Transport

Cite this