The Cells, Vectors, and Protein Expression Core provides advice and support to members ofthe PPG. The Core maintains several suites of vectors used by PPG members for transient, stable and BacMam expression systems in mammalian cells. These include special leader sequences, tag sequences, ligation independent cloning sites, and selectable markers. The Core also supports two types of expression systems for insect cells. The Core maintains cell lines for protein expression, including those with more homogenous glycoforms. It has expertise on growing and cloning hybridoma cell lines. It distributes these materials and provides advice on their use. It advises on methods of protein production, and has facilities for large scale growth of cells for production of recombinant proteins and antibodies. More specifically, the Core provides culture supernatants containing integrin headpiece fragments, protein ligands, and antibodies for Projects 1 and 2, and ICAM-1 D3 and Del-1 fragments for Project 3. It provides baculovirus-infected cells containing FAK and Ser fragments for Project 6. The core will explore production of materials using Pichia for Project 5..

National Institute of Health (NIH)
Research Program Projects (P01)
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Heart, Lung, and Blood Program Project Review Committee (HLBP)
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Children's Hospital Boston
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Springer, Timothy A (2014) von Willebrand factor, Jedi knight of the bloodstream. Blood 124:1412-25
Liu, Ying; Zhang, Yan; Wang, Jia-Huai (2014) Crystal structure of human Ankyrin G death domain. Proteins 82:3476-82
Blenner, Mark A; Dong, Xianchi; Springer, Timothy A (2014) Structural basis of regulation of von Willebrand factor binding to glycoprotein Ib. J Biol Chem 289:5565-79
OuYang, Bo; Chou, James J (2014) The minimalist architectures of viroporins and their therapeutic implications. Biochim Biophys Acta 1838:1058-67
Yu, Yamei; Schurpf, Thomas; Springer, Timothy A (2013) How natalizumab binds and antagonizes *4 integrins. J Biol Chem 288:32314-25
Yu, Yamei; Zhu, Jianghai; Huang, Po-Ssu et al. (2013) Domain 1 of mucosal addressin cell adhesion molecule has an I1-set fold and a flexible integrin-binding loop. J Biol Chem 288:6284-94
Zhu, Jieqing; Zhu, Jianghai; Springer, Timothy A (2013) Complete integrin headpiece opening in eight steps. J Cell Biol 201:1053-68
Wang, Rui; Zhu, Jianghai; Dong, Xianchi et al. (2012) GARP regulates the bioavailability and activation of TGFýý. Mol Biol Cell 23:1129-39
Eng, Edward T; Smagghe, Benoit J; Walz, Thomas et al. (2011) Intact alphaIIbbeta3 integrin is extended after activation as measured by solution X-ray scattering and electron microscopy. J Biol Chem 286:35218-26