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Assay development for lysyl hydroxylase
| Title: | Assay development for lysyl hydroxylase. |
 108 views 24 downloads |
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| Name(s): |
Patel, Deepak A. Florida Atlantic University, Degree grantor Fields, Gregg B., Thesis advisor |
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| Type of Resource: | text | |
| Genre: | Electronic Thesis Or Dissertation | |
| Issuance: | monographic | |
| Date Issued: | 2006 | |
| Publisher: | Florida Atlantic University | |
| Place of Publication: | Boca Raton, Fla. | |
| Physical Form: | application/pdf | |
| Extent: | 67 p. | |
| Language(s): | English | |
| Summary: | Hydroxylysine is produced as a posttranslational modification mainly in collagens, the most abundant protein in mammals. Lysyl hydroxylase (LH) is the enzyme that catalyzes the formation of hydroxylysyl residues in collagen by hydroxylation of -X-Lys-Gly- sequences, for which it requires Fe 2+, 2-oxoglutarate, O2 and ascorbate. In order to study the hydroxylation reaction catalysed by LH, we have synthesized 4 different peptides [for example, GFP*GLP*GAKGE (P*=hydroxyproline) and the corresponding hydroxylated (hydroxylysine-containing) peptide] using Fmoc solid-phase methodology. Peptides have been characterized by HPLC, MALDI-TOF mass spectrometry and CD spectroscopy. A new method for efficient separation of lysine- from hydroxylysine-containing peptides by HPLC has been developed in both organic phase (1-anthroylnitrile as derivatizating reagent) and aqueous phase (dansyl chloride as derivatizating reagent). These reagents have been used to derivatize peptides prior to HPLC analysis. The products (di- and tetra-substituted lysine- and hydroxylysine-containing peptides) have been fully separated by HPLC and their structure confirmed by MALDI-TOF MS analysis. Efficient separation of derivatized peptides will allow for the convenient and rapid measurement of LH activity by HPLC methods. | |
| Identifier: | 9780542745942 (isbn), 13384 (digitool), FADT13384 (IID), fau:10234 (fedora) | |
| Note(s): |
Charles E. Schmidt College of Science Thesis (M.S.)--Florida Atlantic University, 2006. |
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| Subject(s): |
Biological transport Proteins--Metabolism Peptides--Analysis Coenzymes Bioorganic chemistry |
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| Held by: | Florida Atlantic University Libraries | |
| Persistent Link to This Record: | http://purl.flvc.org/fcla/dt/13384 | |
| Sublocation: | Digital Library | |
| Use and Reproduction: | Copyright © is held by the author with permission granted to Florida Atlantic University to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder. | |
| Use and Reproduction: | http://rightsstatements.org/vocab/InC/1.0/ | |
| Host Institution: | FAU | |
| Is Part of Series: | Florida Atlantic University Digital Library Collections. |
