Direct profiling of multiple enzyme activities in human cell lysates by affinity chromatography/electrospray ionization mass spectrometry: application to clinical enzymology

Analytical Chemistry
S A GerberMichael H Gelb

Abstract

We describe a new method for enzyme analysis using affinity capture followed by electrospray ionization mass spectrometry (ACESIMS) for the quantitative determination of the initial velocities of four heparin-modifying enzymes. These enzymes, when defective in affected children, lead to the lysosomal storage disease known as Sanfilippo syndrome. The method relies on substrates and internal standards conjugated to the molecular handle biotin via a heavy isotope-encodable, mass-adjustable linker. Reaction velocities of the Sanfilippo enzymes in a crude lysate prepared from as little as 2500 human skin fibroblasts can be determined. In addition, the ACESIMS method is widely applicable to the simultaneous analysis of multiple enzymes in a complex biological sample by a single analytical technique and will thus serve as a useful tool in basic and clinical biomedical research.

References

Jan 1, 1992·Human Mutation·J K Reichardt
Jul 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·J S O'Brien
Nov 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·H KresseW Fuchs
Nov 22, 1994·Proceedings of the National Academy of Sciences of the United States of America·G Siuzdak
Jan 1, 1996·Journal of Inherited Metabolic Disease·E A KarpovaO P van Diggelen
Feb 27, 1999·Current Opinion in Biotechnology·C E Costello
Jan 5, 2000·Proceedings of the National Academy of Sciences of the United States of America·R SchiffmannR O Brady
Jan 8, 2000·Acta Paediatrica. Supplement·E R McCabe, L L McCabe
Jun 29, 2000·Annual Review of Biochemistry·M BernfieldM Zako

❮ Previous
Next ❯

Citations

Jul 12, 2005·Analytical and Bioanalytical Chemistry·André Liesener, Uwe Karst
Jul 20, 2011·Analytical and Bioanalytical Chemistry·Johanna GrassmannThomas Letzel
May 26, 2005·Journal of the American Society for Mass Spectrometry·Huili ZhaiFred W McLafferty
Feb 5, 2005·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Ed J WraithDoug A Brooks
Feb 12, 2011·American Journal of Medical Genetics. Part C, Seminars in Medical Genetics·Kimitoshi NakamuraFumio Endo
Feb 11, 2005·Current Opinion in Chemical Biology·Nicola L Pohl
Apr 16, 2011·The Journal of Pediatrics·Hui ZhouRobert F Vogt
Dec 4, 2014·Analytica Chimica Acta·Radoslaw KwapiszewskiZbigniew Brzozka
Sep 7, 2004·Expert Review of Molecular Diagnostics·Peter J MeikleJohn J Hopwood
Jan 1, 2012·European Journal of Mass Spectrometry·Tingting CaiYinlong Guo
Aug 28, 2001·Journal of Mass Spectrometry : JMS
May 22, 2010·Clinical Chemistry·Deborah Marsden, Harvey Levy
Apr 28, 2005·Clinical Chemistry·David S Millington
Sep 25, 2004·Clinical Chemistry·Peter Clements
Apr 12, 2020·Journal of Human Genetics·Nivethitha ArunkumarShunji Tomatsu
Apr 17, 2013·Analytical Chemistry·Barbara GregoriusAndreas Tholey
Jan 25, 2002·Analytical Chemistry·Michele A KellyPhilip J Rosner
Jan 22, 2004·Bioconjugate Chemistry·Anupama Patel, David M Perrin

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.