fulltext.study @t Gmail

Synthesis of substrates for periodate-coupled assay of phospholipases C and sphingomyelinases

Paper ID Volume ID Publish Year Pages File Format Full-Text
16752 42609 2016 6 PDF Available
Title
Synthesis of substrates for periodate-coupled assay of phospholipases C and sphingomyelinases
Abstract

•Four substrate analogues of glycerophospholipids are synthesized.•One substrate is evaluated in periodate-coupled assays of fungal sphingomyelinases.•Periodate-containing stop solutions are optimized.•A kinetic evaluation showed relatively high KM and low kcat values.

A series of 4-nitrophenyl (pNP) and 4-methylumbelliferyl (4MU) substrate analogues of phosphatidyl choline (PC) and phosphatidic acid (PA) were synthesized from 4-bromo-1-butene by ether formation, olefin epoxidation and ring opening with the phosphate head group. The pNP PC analogue, 4-(4-nitrophenoxy)-2-hydroxy-butyl-1-phosphoryl choline (1) was evaluated in assays of fungal sphingomyelinases, also displaying phospholipase C activity. Reactions were terminated with a periodate-containing stop solution, leading to liberation of pNP, quantified spectrophotometrically in an end-point measurement. A kinetic evaluation of sphingomyelinases from Kionochaeta sp. and Penicillium emersonii showed relatively high KM and low kcat values for this substrate, limiting its practical applicability in assays with low sphingomyelinase concentrations.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slide

Keywords
Periodate-coupled assay; Phospholipase C (PLC); Sphingomyelinase; Synthetic assay substrate; Enzyme kinetics
First Page Preview
Synthesis of substrates for periodate-coupled assay of phospholipases C and sphingomyelinases
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 91, September 2016, Pages 66–71
Authors
, , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us