SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIES
The first example of a kinetic labeling library designed to enable the discovery of affinity labels is presented. Each library component (1) consists of a variable peptidyl component linked to a biotinyl moiety by a 4-mercaptobenzoyl linker in thioester format. We demonstrate that an affinity label...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2014-03-01
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| Series: | Computational and Structural Biotechnology Journal |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2001037014600015 |
| _version_ | 1828832494452998144 |
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| author | Allen Krantz Arthur M Hanel Ivona Strug Andrzej Wilczynski Jeremy J Wolff Wolin Huang Linda H Huang Tina Settineri Darren L Holmes Margaret C Hardy Dominique P Bridon |
| author_facet | Allen Krantz Arthur M Hanel Ivona Strug Andrzej Wilczynski Jeremy J Wolff Wolin Huang Linda H Huang Tina Settineri Darren L Holmes Margaret C Hardy Dominique P Bridon |
| author_sort | Allen Krantz |
| collection | DOAJ |
| description | The first example of a kinetic labeling library designed to enable the discovery of affinity labels is presented. Each library component (1) consists of a variable peptidyl component linked to a biotinyl moiety by a 4-mercaptobenzoyl linker in thioester format. We demonstrate that an affinity label can be uncovered by measuring reaction rates between library pools and the protein target, human serum albumin (HSA) and identifying significant outliers. By choosing peptide functionality compatible with a potentially reactive thioester labeling entity, libraries can be screened in pools. It is noteworthy that a limited subset of amino acids (R, S, E, F, Y, l, M, W, and Q) that compose the affinity moiety is sufficient to produce rate variances that guide the discovery process. After two rounds of deconvolution, J-FLYEE-NH2 (7-E) emerges as a bona fide affinity label of HSA. Unlike known affinity labels, the affinity moiety is not retained in the protein product, but is extruded upon acylation of the protein. This feature affords a method of introducing various payloads, without extraneous elements, onto protein frameworks. |
| first_indexed | 2024-12-12T16:55:14Z |
| format | Article |
| id | doaj.art-b5781b56027b45358e4cbb98b9eca02e |
| institution | Directory Open Access Journal |
| issn | 2001-0370 |
| language | English |
| last_indexed | 2024-12-12T16:55:14Z |
| publishDate | 2014-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Computational and Structural Biotechnology Journal |
| spelling | doaj.art-b5781b56027b45358e4cbb98b9eca02e2022-12-22T00:18:14ZengElsevierComputational and Structural Biotechnology Journal2001-03702014-03-0191510.5936/csbj.201403001SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIESAllen Krantz0Arthur M Hanel1Ivona Strug2Andrzej Wilczynski3Jeremy J Wolff4Wolin Huang5Linda H Huang6Tina Settineri7Darren L Holmes8Margaret C Hardy9Dominique P Bridon10Advanced Proteome Therapeutics Inc., 650 Albany Street, Suite 113, Boston, MA 02118, United StatesRedCell Inc., 270-B Littlefield Avenue, South San Francisco, CA, 94080, United States (Renamed ConjuChem LLC. Current address: 11755 Wilshire Blvd, Suite 2000, Los Angeles, CA 90025.)Advanced Proteome Therapeutics Inc., 650 Albany Street, Suite 113, Boston, MA 02118, United StatesAdvanced Proteome Therapeutics Inc., 650 Albany Street, Suite 113, Boston, MA 02118, United StatesBruker Daltonics Inc., 40 Manning Road, Billerica, MA 01821, United StatesRedCell Inc., 270-B Littlefield Avenue, South San Francisco, CA, 94080, United States (Renamed ConjuChem LLC. Current address: 11755 Wilshire Blvd, Suite 2000, Los Angeles, CA 90025.)RedCell Inc., 270-B Littlefield Avenue, South San Francisco, CA, 94080, United States (Renamed ConjuChem LLC. Current address: 11755 Wilshire Blvd, Suite 2000, Los Angeles, CA 90025.)RedCell Inc., 270-B Littlefield Avenue, South San Francisco, CA, 94080, United States (Renamed ConjuChem LLC. Current address: 11755 Wilshire Blvd, Suite 2000, Los Angeles, CA 90025.)RedCell Inc., 270-B Littlefield Avenue, South San Francisco, CA, 94080, United States (Renamed ConjuChem LLC. Current address: 11755 Wilshire Blvd, Suite 2000, Los Angeles, CA 90025.)RedCell Inc., 270-B Littlefield Avenue, South San Francisco, CA, 94080, United States (Renamed ConjuChem LLC. Current address: 11755 Wilshire Blvd, Suite 2000, Los Angeles, CA 90025.)RedCell Inc., 270-B Littlefield Avenue, South San Francisco, CA, 94080, United States (Renamed ConjuChem LLC. Current address: 11755 Wilshire Blvd, Suite 2000, Los Angeles, CA 90025.)The first example of a kinetic labeling library designed to enable the discovery of affinity labels is presented. Each library component (1) consists of a variable peptidyl component linked to a biotinyl moiety by a 4-mercaptobenzoyl linker in thioester format. We demonstrate that an affinity label can be uncovered by measuring reaction rates between library pools and the protein target, human serum albumin (HSA) and identifying significant outliers. By choosing peptide functionality compatible with a potentially reactive thioester labeling entity, libraries can be screened in pools. It is noteworthy that a limited subset of amino acids (R, S, E, F, Y, l, M, W, and Q) that compose the affinity moiety is sufficient to produce rate variances that guide the discovery process. After two rounds of deconvolution, J-FLYEE-NH2 (7-E) emerges as a bona fide affinity label of HSA. Unlike known affinity labels, the affinity moiety is not retained in the protein product, but is extruded upon acylation of the protein. This feature affords a method of introducing various payloads, without extraneous elements, onto protein frameworks.http://www.sciencedirect.com/science/article/pii/S2001037014600015Site-specific protein modificationKinetic labeling librariesAffinity labelsLysine labelingCombinatorial librariesPeptide libraries |
| spellingShingle | Allen Krantz Arthur M Hanel Ivona Strug Andrzej Wilczynski Jeremy J Wolff Wolin Huang Linda H Huang Tina Settineri Darren L Holmes Margaret C Hardy Dominique P Bridon SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIES Computational and Structural Biotechnology Journal Site-specific protein modification Kinetic labeling libraries Affinity labels Lysine labeling Combinatorial libraries Peptide libraries |
| title | SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIES |
| title_full | SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIES |
| title_fullStr | SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIES |
| title_full_unstemmed | SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIES |
| title_short | SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIES |
| title_sort | site specific labeling of a protein lysine residue by novel kinetic labeling combinatorial libraries |
| topic | Site-specific protein modification Kinetic labeling libraries Affinity labels Lysine labeling Combinatorial libraries Peptide libraries |
| url | http://www.sciencedirect.com/science/article/pii/S2001037014600015 |
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