PUBLICATION
A High-Throughput Colorimetric Assay to Characterize the Enzyme Kinetic and Cellular Activity of Spermidine/Spermine N1-Acetyltransferase 1
- Authors
- Lin, H.J., Lien, Y.C., and Hsu, C.H.
- ID
- ZDB-PUB-100811-7
- Date
- 2010
- Source
- Analytical biochemistry 407(2): 226-232 (Journal)
- Registered Authors
- Keywords
- Activity assay, Ellman’s reagent, Enzyme kinetics, Polyamines, Spermidine/spermine N1-acetyltransferase 1
- MeSH Terms
-
- Acetyltransferases/chemistry
- Acetyltransferases/genetics
- Acetyltransferases/metabolism*
- Animals
- Colorimetry/methods*
- Enzyme Assays/methods*
- High-Throughput Screening Assays/methods*
- Kinetics
- Polyamines/metabolism
- Recombinant Proteins/chemistry
- Recombinant Proteins/genetics
- Recombinant Proteins/metabolism
- Zebrafish
- Zebrafish Proteins/chemistry
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- PubMed
- 20692222 Full text @ Anal. Biochem.
Citation
Lin, H.J., Lien, Y.C., and Hsu, C.H. (2010) A High-Throughput Colorimetric Assay to Characterize the Enzyme Kinetic and Cellular Activity of Spermidine/Spermine N1-Acetyltransferase 1. Analytical biochemistry. 407(2):226-232.
Abstract
Spermidine/spermine N1-acetyltransferase 1 (SSAT1) is a key enzyme that catalyzes the catabolism of polyamines. SSAT1 is a very important enzyme because it not only maintains the homeostasis of polyamines but is also involved in many physiological and pathological events. As such, a rapid assay of SSAT1 activity is valuable in drug screening and clinical diagnostics. Here, we report a novel colorimetric assay for monitoring SSAT1 activity in zebrafish (zSSAT1). In comparison with the available SSAT1 assays, this new method is cost-effective, and simple. The optimal zSSAT1 activity was obtained below 55 degrees C in a mild alkaline environment. The Km of zSSAT1 for spermidine and spermine is 55 and 182 muM, respectively, while putrescine is not a good substrate for zSSAT1. In addition to enzyme kinetic studies, the colorimetric assay was also used to detect the cellular activity of SSAT1. Thus, the present method is a reliable assay for determining SSAT1 activity with many potential applications in medical biology.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping