Tandem Mass Spectrometry Assays of Palmitoyl Protein Thioesterase 1 and Tripeptidyl Peptidase Activity in Dried Blood Spots for the Detection of Neuronal Ceroid Lipofuscinoses in Newborns

被引：17

作者：

Barcenas, Mariana ^{[1
]}

Xue, Chang ^{[1
]}

Marushchak-Vlaskin, Tatyana ^{[1
]}

Scott, C. Ronald ^{[2
]}

Gelb, Michael H. ^{[1
,3
]}

Turecek, Frantisek ^{[1
]}

机构：

[1] Univ Washington, Dept Chem, Seattle, WA 98115 USA

[2] Univ Washington, Dept Pediat, Seattle, WA 98115 USA

[3] Univ Washington, Dept Biochem, Seattle, WA 98115 USA

来源：

ANALYTICAL CHEMISTRY | 2014年 / 86卷 / 15期

关键词：

LYSOSOMAL-ENZYMES; DIAGNOSIS;

D O I：

10.1021/ac501994b

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

We report new substrates for quantitative enzyme activity measurements of human palmitoyl protein thioesterase (PPT1) and tripeptidyl peptidase (TPP1) in dried blood spots from newborns using tandem mass spectrometry. Deficiencies in these enzyme activities due to inborn errors of metabolism cause neuronal ceroid lipofuscinoses. The assays use synthetic compounds that were designed to mimic the natural substrates. Incubation produces nanomole quantities of enzymatic products per a blood spot that are quantified by tandem mass spectrometry using synthetic internal standards and selected reaction monitoring. The assays utilize a minimum steps for sample workup and can be run in a duplex format for the detection of neuronal ceroid lipofuscinoses or potentially multiplexed with other mass spectrometry-based assays for newborn screening of lysosomal storage disorders.

引用

页码：7962 / 7968

页数：7

共 25 条

[1] The crystal structure of palmitoyl protein thioesterase 1 and the molecular basis of infantile neuronal ceroid lipofuscinosis [J].