Deepak Patel

Broschiertes Buch

Assay Development for Lysyl Hydroxylase

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

Beschreibung

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.

Autor

Deepak A. Patel was born in Surat, India, on March 20, 1977. He graduated from Mumbai University, India in 1998 with a degree in Chemistry. For several years he worked as a chemist in Mumbai, India, before relocating to Florida, USA, to pursue higher study in Analytical Chemistry. He completed Master of Science in 2006.

Produktdetails

• Verlag: VDM Verlag Dr. Müller
• Seitenzahl: 68
• Englisch
• ISBN-13: 9783639222579
• ISBN-10: 3639222571
• Artikelnr.: 28253718