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Investigating the Dynamics and Polyanion Binding Sites of Fibroblast Growth Factor-1 Using Hydrogen-Deuterium Exchange Mass Spectrometry

Title: Investigating the Dynamics and Polyanion Binding Sites of Fibroblast Growth Factor-1 Using Hydrogen-Deuterium Exchange Mass Spectrometry.
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Name(s): Angalakurthi, Siva K, author
Tenorio, Connie A, author
Blaber, Michael, author
Middaugh, Russell, author
Type of Resource: text
Genre: Text
Working Paper
Date Issued: 2018-04-05
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: In this study, we examined the local dynamics of acidic fibroblast growth factor (FGF-1) as well as the binding sites of various polyanions including poly-sulfates (heparin and low MW heparin) and poly-phosphates (phytic acid and ATP) using hydrogen-deuterium exchange mass spectrometry (HX-MS). For local dynamics, results are analyzed at the peptide level as well as in terms of buried amides employing crystallographic B-factors and compared with a residue level heat map generated from HX-MS results. Results show that strand 4 and 5 and the turn between them to be the most flexible regions as was previously seen by NMR. On the other hand, the C-terminal strands 8, 9 and 10 appear to be more rigid which is also consistent with crystallographic B-factors as well as local dynamics studies conducted by NMR. Crystal structures of FGF-1 in complex with heparin have shown that heparin binds to N-terminal Asn18 and to C-terminal Lys105, Tryp107, Lys112, Lys113, Arg119, Pro121, Arg122, Gln127 and Lys128 indicating electrostatic forces as dominant interactions. Heparin binding as determined by HX-MS is consistent with crystallography data. Previous studies have also shown that other polyanions including low MW heparin, phytic acid and ATP dramatically increase the thermal stability of FGF-1. Using HX-MS, we find other poly anions tested bind in a similar manner to heparin, primarily targeting the turns in the lysine rich C-terminal region of FGF-1 along with two distinct N-terminal regions that contains lysines and arginines/ histidines. This confirms the interactions between FGF-1 and polyanions are primary directed by electrostatics.
Identifier: FSU_libsubv1_scholarship_submission_1523040928_8f170751 (IID)
Keywords: Ligand binding, protein backbone dynamics, polyanions, charge mediated interactions, thermal stability, HX-MS
Publication Note: This is the pre-peer reviewed version of the following article: Investigating the Dynamics and Polyanion Binding Sites of Fibroblast Growth Factor-1 Using Hydrogen-Deuterium Exchange Mass Spectrometry, which has been published in final form in Protein Science. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_libsubv1_scholarship_submission_1523040928_8f170751
Owner Institution: FSU
Is Part Of: Protein Science.

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Angalakurthi, S. K., Tenorio, C. A., Blaber, M., & Middaugh, R. (2018). Investigating the Dynamics and Polyanion Binding Sites of Fibroblast Growth Factor-1 Using Hydrogen-Deuterium Exchange Mass Spectrometry. Protein Science. Retrieved from http://purl.flvc.org/fsu/fd/FSU_libsubv1_scholarship_submission_1523040928_8f170751