Microarray of human P450 with an integrated oxygen sensing film for high-throughput detection of metabolic activities

Analytical Chemistry
Gang ChangKenichi Morigaki

Abstract

A microarray chip containing human P450 isoforms was constructed for the parallel assay of their metabolic activities. The chip had microwells that contained vertically integrated P450 and oxygen sensing layers. The oxygen sensing film was made of an organically modified silica film (ORMOSIL) doped with tris(4,7-diphenyl-1,10-phenanthroline) ruthenium dichloride (Ru(dpp)(3)Cl(2)). Human P450s (23 types) expressed in E. coli and purified as membrane fractions were immobilized in agarose matrixes on the oxygen sensing layer. The activities of P450s were determined by evaluating the fluorescence intensity enhancement of the oxygen sensor due to the oxygen consumption by the metabolic reaction. By normalizing the responses with the amounts of oxygen sensor and P450 enzymes in microwells, we could obtain fluorescence enhancement patterns that were characteristic to the combination of P450 isoforms and substrate material. The patterns obtained from two psoralen derivatives resembled each other, whereas a structurally different substrate (capsaicin) resulted in a distinct pattern. These results suggest the potential of the microarray to analyze the activities of diverse P450 isoforms in a high-throughput fashion. Furthermore, mechanis...Continue Reading

References

Jul 17, 1998·Biochemistry·L L Koenigs, W F Trager
Dec 18, 2001·Biosensors & Bioelectronics·Georgia Eleni TsotsouGianfranco Gilardi
Jan 31, 2003·Biotechnology and Bioengineering·Gernot T JohnElmar Heinzle
Jan 6, 2004·Journal of Pharmaceutical Sciences·John H Ansede, Dhiren R Thakker
Jan 20, 2005·Proceedings of the National Academy of Sciences of the United States of America·Moo-Yeal LeeDouglas S Clark
Apr 28, 2005·Biosensors & Bioelectronics·Nikitas BistolasFrieder W Scheller
Nov 1, 2005·Analytical Chemistry·Kumiko Sakai-KatoNaoko Utsunomiya-Tate
May 27, 2006·Current Drug Metabolism·Chul-Ho YunTaeho Ahn
Jul 7, 2007·Environmental Science & Technology·Hei-Leung PangKwok-Yin Wong
Dec 28, 2007·Proceedings of the National Academy of Sciences of the United States of America·Moo-Yeal LeeJonathan S Dordick
Jan 31, 2008·Chemical Reviews·Sergey M Borisov, Otto S Wolfbeis
Nov 11, 2008·Current Drug Metabolism·Robert L Walsky, Sherri E Boldt
Jun 12, 2009·Drug Metabolism Reviews·Shu-Feng ZhouBalram Chowbay
Aug 28, 2010·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Tatsushi GotoHiromasa Imaishi
Jul 20, 2011·Journal of the American Chemical Society·Matthew J TraylorDouglas S Clark

❮ Previous
Next ❯

Citations


❮ 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.