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insight into the thermodynamic characteristics of human thrombopoietin complexation with TN1 antibody.

Title: An insight into the thermodynamic characteristics of human thrombopoietin complexation with TN1 antibody.
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Name(s): Arai, Shigeki, author
Shibazaki, Chie, author
Adachi, Motoyasu, author
Honjo, Eijiro, author
Tamada, Taro, author
Maeda, Yoshitake, author
Tahara, Tomoyuki, author
Kato, Takashi, author
Miyazaki, Hiroshi, author
Blaber, Michael, author
Kuroki, Ryota, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2016-10-01
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Human thrombopoietin (hTPO) primarily stimulates megakaryocytopoiesis and platelet production and is neutralized by the mouse TN1 antibody. The thermodynamic characteristics of TN1 antibody-hTPO complexation were analyzed by isothermal titration calorimetry (ITC) using an antigen-binding fragment (Fab) derived from the TN1 antibody (TN1-Fab). To clarify the mechanism by which hTPO is recognized by TN1-Fab the conformation of free TN1-Fab was determined to a resolution of 2.0 Å using X-ray crystallography and compared with the hTPO-bound form of TN1-Fab determined by a previous study. This structural comparison revealed that the conformation of TN1-Fab does not substantially change after hTPO binding and a set of 15 water molecules is released from the antigen-binding site (paratope) of TN1-Fab upon hTPO complexation. Interestingly, the heat capacity change (ΔCp) measured by ITC (-1.52 ± 0.05 kJ mol(-1)  K(-1) ) differed significantly from calculations based upon the X-ray structure data of the hTPO-bound and unbound forms of TN1-Fab (-1.02 ∼ 0.25 kJ mol(-1)  K(-1) ) suggesting that hTPO undergoes an induced-fit conformational change combined with significant desolvation upon TN1-Fab binding. The results shed light on the structural biology associated with neutralizing antibody recognition.
Identifier: FSU_pmch_27419667 (IID), 10.1002/pro.2985 (DOI), PMC5029525 (PMCID), 27419667 (RID), 27419667 (EID)
Keywords: TN1, X-ray crystallography, Antigen-antibody interaction, Isothermal titration calorimetry, Thrombopoietin
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5029525.
Subject(s): Animals
Antibodies, Monoclonal, Murine-Derived/chemistry
Crystallography, X-Ray
Humans
Immunoglobulin Fab Fragments/chemistry
Mice
Protein Structure, Quaternary
Thermodynamics
Thrombopoietin/chemistry
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_27419667
Owner Institution: FSU
Is Part Of: Protein science : a publication of the Protein Society.
1469-896X
Issue: iss. 10, vol. 25

Choose the citation style.
Arai, S., Shibazaki, C., Adachi, M., Honjo, E., Tamada, T., Maeda, Y., … Kuroki, R. (2016). An insight into the thermodynamic characteristics of human thrombopoietin complexation with TN1 antibody. Protein Science : A Publication Of The Protein Society. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_27419667