This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Beside hemostasis, platelets are known to play a role in immune responses and inflammatory processes. Interaction with pathogens involves pathogen recognition, release of granule-content, and cytoskeletal rearrangements in the platelet. We have identified the expression of toll-like receptors (TLR) on the platelet surface, supporting a role for the platelet in innate immune responses. In addition, the TLR pathway links inflammation to atherosclerosis, as TLRs are expressed in atherosclerotic plaques and murine TLR2 stimulation causes enhanced atherosclerosis. Intracellular Factor XIIIA (FXIIIA) can crosslink platelet cytoskeletal proteins upon platelet activation. Relevant to the immune response, FXIIIA is required for cytoskeletal remodeling during monocyte phagocytosis, but its role in platelet-mediated immune processes is unknown. We have applied a proteomics approach to search for TLR2- and FXIIIA-associated proteins in resting platelets. After immunoprecipitation (IP) of TLR2 and FXIIIA from platelet lysates followed by gel separation, protein bands were excised, digested with trypsin and analysed by mass spectrometry. Western blot analysis was applied to confirm the mass spectrometric results. We repeatedly immunoprecipitated FXIIIA directly with TLR2-specific antibodies, whereas the IP of FXIIIA pulled down a truncated form of TLR2, as identified by Western blot, that may correspond to the soluble form of TLR2. By mass spectrometry, we identified FXIIIA and FXIIIA-associated proteins: thrombospondin, filamin and talin. Importantly, talin has not been previously reported as FXIII-substrate or associated protein. Thrombospondin, a known FXIII substrate, is stored in platelet granules. A strong thrombospondin-band after FXIIIA-IP and a weak band after TLR2-IP suggests the partial presence of FXIIIA and soluble TLR2 in granules. Interestingly, FXIIIA appears to be associated with its substrates in the resting platelet. In conclusion, our results indicate the presence of known and novel FXIIIA-associated proteins in platelets and suggests a role for TLR2 and FXIIIA in platelet-dependent immune responses. A manuscript reporting these results has been accepted for publication in Thrombosis and Hemostasis .

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR010888-13
Application #
7955929
Study Section
Special Emphasis Panel (ZRG1-BCMB-H (40))
Project Start
2009-06-01
Project End
2010-05-31
Budget Start
2009-06-01
Budget End
2010-05-31
Support Year
13
Fiscal Year
2009
Total Cost
$2,344
Indirect Cost
Name
Boston University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02118
Lu, Yanyan; Jiang, Yan; Prokaeva, Tatiana et al. (2017) Oxidative Post-Translational Modifications of an Amyloidogenic Immunoglobulin Light Chain Protein. Int J Mass Spectrom 416:71-79
Sethi, Manveen K; Zaia, Joseph (2017) Extracellular matrix proteomics in schizophrenia and Alzheimer's disease. Anal Bioanal Chem 409:379-394
Hu, Han; Khatri, Kshitij; Zaia, Joseph (2017) Algorithms and design strategies towards automated glycoproteomics analysis. Mass Spectrom Rev 36:475-498
Ji, Yuhuan; Bachschmid, Markus M; Costello, Catherine E et al. (2016) S- to N-Palmitoyl Transfer During Proteomic Sample Preparation. J Am Soc Mass Spectrom 27:677-85
Hu, Han; Khatri, Kshitij; Klein, Joshua et al. (2016) A review of methods for interpretation of glycopeptide tandem mass spectral data. Glycoconj J 33:285-96
Pu, Yi; Ridgeway, Mark E; Glaskin, Rebecca S et al. (2016) Separation and Identification of Isomeric Glycans by Selected Accumulation-Trapped Ion Mobility Spectrometry-Electron Activated Dissociation Tandem Mass Spectrometry. Anal Chem 88:3440-3
Wang, Yun Hwa Walter; Meyer, Rosana D; Bondzie, Philip A et al. (2016) IGPR-1 Is Required for Endothelial Cell-Cell Adhesion and Barrier Function. J Mol Biol 428:5019-5033
Srinivasan, Srimathi; Chitalia, Vipul; Meyer, Rosana D et al. (2015) Hypoxia-induced expression of phosducin-like 3 regulates expression of VEGFR-2 and promotes angiogenesis. Angiogenesis 18:449-62
Yu, Xiang; Sargaeva, Nadezda P; Thompson, Christopher J et al. (2015) In-Source Decay Characterization of Isoaspartate and ?-Peptides. Int J Mass Spectrom 390:101-109
Steinhorn, Benjamin S; Loscalzo, Joseph; Michel, Thomas (2015) Nitroglycerin and Nitric Oxide--A Rondo of Themes in Cardiovascular Therapeutics. N Engl J Med 373:277-80

Showing the most recent 10 out of 253 publications