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Assay Development for Lysyl Hydroxylase

Name: Assay Development for Lysyl Hydroxylase
Brand: VDM Verlag Dr. Müller
SKU: 06830083
Price: 41.59 EUR
Availability: InStock
Author: Deepak Patel
ISBN: 9783639222579

A new HPLC method for efficient separation of lysine- from hydroxylysine-containing peptides

Deepak Patel

Lingua

Inglese

Libro In brossura

Codice Libristo: 06830083

Casa editrice VDM Verlag Dr. Müller, novembre 2009

Hydroxylysine is produced as a posttranslational modification mainly in collagens, the most abundant... Descrizione completa

Codice Libristo: 06830083

102 b

41.59 €

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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. In order to study the hydroxylation reaction catalysed by LH, we have synthesized 4 different peptides and the corresponding hydroxylated peptide using Fmoc solid-phase methodology. Peptides have been characterized by HPLC, mass spectrometry and CD spectroscopy. A new HPLC method for efficient separation of lysine- from hydroxylysine-containing peptides has been developed in both organic (1-anthroylnitrile as derivatizating reagent) and aqueous phase (dansyl chloride as derivatizating reagent). Peptides derivatization is done 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.