M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins
We exploit bacterial sortases to attach a variety of moieties to the capsid proteins of M13 bacteriophage. We show that pIII, pIX, and pVIII can be functionalized with entities ranging from small molecules (e.g., fluorophores, biotin) to correctly folded proteins (e.g., GFP, antibodies, streptavidin...
| Main Authors: | , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
American Chemical Society (ACS)
2013
|
| Online Access: | http://hdl.handle.net/1721.1/80400 https://orcid.org/0000-0001-9353-7453 https://orcid.org/0000-0002-1090-6071 |
| _version_ | 1826201583652700160 |
|---|---|
| author | Hess, Gaelen T. Cragnolini, Juan J. Popp, Maximilian W. Allen, Mark A. Dougan, Stephanie K. Spooner, Eric Ploegh, Hidde L. Belcher, Angela M. Guimaraes, Carla P. |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Hess, Gaelen T. Cragnolini, Juan J. Popp, Maximilian W. Allen, Mark A. Dougan, Stephanie K. Spooner, Eric Ploegh, Hidde L. Belcher, Angela M. Guimaraes, Carla P. |
| author_sort | Hess, Gaelen T. |
| collection | MIT |
| description | We exploit bacterial sortases to attach a variety of moieties to the capsid proteins of M13 bacteriophage. We show that pIII, pIX, and pVIII can be functionalized with entities ranging from small molecules (e.g., fluorophores, biotin) to correctly folded proteins (e.g., GFP, antibodies, streptavidin) in a site-specific manner, and with yields that surpass those of any reported using phage display technology. A case in point is modification of pVIII. While a phage vector limits the size of the insert into pVIII to a few amino acids, a phagemid system limits the number of copies actually displayed at the surface of M13. Using sortase-based reactions, a 100-fold increase in the efficiency of display of GFP onto pVIII is achieved. Taking advantage of orthogonal sortases, we can simultaneously target two distinct capsid proteins in the same phage particle and maintain excellent specificity of labeling. As demonstrated in this work, this is a simple and effective method for creating a variety of structures, thus expanding the use of M13 for materials science applications and as a biological tool. |
| first_indexed | 2024-09-23T11:53:43Z |
| format | Article |
| id | mit-1721.1/80400 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:53:43Z |
| publishDate | 2013 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/804002022-10-01T06:47:08Z M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins Hess, Gaelen T. Cragnolini, Juan J. Popp, Maximilian W. Allen, Mark A. Dougan, Stephanie K. Spooner, Eric Ploegh, Hidde L. Belcher, Angela M. Guimaraes, Carla P. Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Materials Science and Engineering Koch Institute for Integrative Cancer Research at MIT Ploegh, Hidde Hess, Gaelen T. Allen, Mark A. Belcher, Angela M. Ploegh, Hidde We exploit bacterial sortases to attach a variety of moieties to the capsid proteins of M13 bacteriophage. We show that pIII, pIX, and pVIII can be functionalized with entities ranging from small molecules (e.g., fluorophores, biotin) to correctly folded proteins (e.g., GFP, antibodies, streptavidin) in a site-specific manner, and with yields that surpass those of any reported using phage display technology. A case in point is modification of pVIII. While a phage vector limits the size of the insert into pVIII to a few amino acids, a phagemid system limits the number of copies actually displayed at the surface of M13. Using sortase-based reactions, a 100-fold increase in the efficiency of display of GFP onto pVIII is achieved. Taking advantage of orthogonal sortases, we can simultaneously target two distinct capsid proteins in the same phage particle and maintain excellent specificity of labeling. As demonstrated in this work, this is a simple and effective method for creating a variety of structures, thus expanding the use of M13 for materials science applications and as a biological tool. United States. Army Research Office. (Institute for Collaborative Biotechnologies Grant W911NF-09-0001) 2013-09-11T18:36:12Z 2013-09-11T18:36:12Z 2012-07 2012-06 Article http://purl.org/eprint/type/JournalArticle 1043-1802 1520-4812 http://hdl.handle.net/1721.1/80400 Hess, Gaelen T., Juan J. Cragnolini, Maximilian W. Popp, Mark A. Allen, Stephanie K. Dougan, Eric Spooner, Hidde L. Ploegh, Angela M. Belcher, and Carla P. Guimaraes. “M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins.” Bioconjugate Chemistry 23, no. 7 (July 18, 2012): 1478-1487. https://orcid.org/0000-0001-9353-7453 https://orcid.org/0000-0002-1090-6071 en_US http://dx.doi.org/10.1021/bc300130z Bioconjugate Chemistry Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society (ACS) Ploegh via Courtney Crummett |
| spellingShingle | Hess, Gaelen T. Cragnolini, Juan J. Popp, Maximilian W. Allen, Mark A. Dougan, Stephanie K. Spooner, Eric Ploegh, Hidde L. Belcher, Angela M. Guimaraes, Carla P. M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins |
| title | M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins |
| title_full | M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins |
| title_fullStr | M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins |
| title_full_unstemmed | M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins |
| title_short | M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins |
| title_sort | m13 bacteriophage display framework that allows sortase mediated modification of surface accessible phage proteins |
| url | http://hdl.handle.net/1721.1/80400 https://orcid.org/0000-0001-9353-7453 https://orcid.org/0000-0002-1090-6071 |
| work_keys_str_mv | AT hessgaelent m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins AT cragnolinijuanj m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins AT poppmaximilianw m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins AT allenmarka m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins AT douganstephaniek m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins AT spoonereric m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins AT ploeghhiddel m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins AT belcherangelam m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins AT guimaraescarlap m13bacteriophagedisplayframeworkthatallowssortasemediatedmodificationofsurfaceaccessiblephageproteins |