[광원으로 온-오프 가능한 CENP-E 억제제/Photoswitchable CENP-E Inhibitor] PCEI-HU [FDV-0037]_Funakoshi

Controllable by light! Chromosome movement regulation reagent in mitosis

PCEI-HU

Photoswitchable CENP-E Inhibitor

Cat.# FDV-0037 / Size: 50ug

 

PCEI-HU(Photoswitchable CENP-E inhibitor)는 체세포분열(유사분열) 중기에 Centromere protein E의 움직임을 조절할 수 있는 세계 최초 시약 입니다. 가시광은 염색체 이동을 멈추게 하고 자외선은 재개 시킵니다. 이 움직임은 모든 염색체가 중기판(metaphase plate)에 정렬 될 때까지 제어 가능합니다.

 

PCEI-HU는 전중기 / 중기 및 SAC(Spindle Assembly Checkpoint)의 기초 및 핵심 연구를 위한 강력 추천 합니다!

 

PCEI-HU (Photoswitchable CENP-E inhibitor) is the world's first reagent which can control movements of Centromere protein E (CENP-E), a kinesin-like motor protein in metapahse of mitosis. Visible light stops the movement of chromosome, and UV light resumes the movement of chromosome. This movement is controllable until all chromosomes are aligned on the metaphase plate in metaphase.

 

PCEI-HU is a powerful tool for the fundamental investigation of prometaphase/metaphase and spindle assembly checkpoint (SAC).

 

 

This product has been commercialized under the license from Hokkaido University.

 

Product Background 

Mitosis is a fundamental process of cell duplication for the growth and maintenance of tissues in multicellular organisms. During mitosis, many kinds of proteins play crucial roles in accurate distribution of the replicated genome into two daughter cells. The replicated genomes are condensed into chromosomes in the prophase and subsequently, chromosomes move to the center of the mitotic cells, frequently called metaphase plate, along spindle microtubules in prometaphase. Chromosomes bind to spindle microtubules via kinetochore which is a unique protein complex in centromere. Only when all chromosomes are aligned to the metaphase plate in metaphase, separation of sister chromosome begins (anaphase). There is an important checkpoint called spindle assembly checkpoint (SAC) from metaphase to anaphase. SAC monitors accurate alignment of chromosomes in metaphase plate in mitosis to exact separation of genomic information into daughter cells. Accurate segregation of all chromosomes is the most important matter to satisfy the SAC criteria. 

 

Centromere protein E (CENP-E) is a key component for chromosome congression along microtubule spindles to the metaphase plate. CENP-E is a dimeric protein which has three domains, N-terminal motor domain, C-terminal kinetochore binding domain and coiled-coiled linker region. Recent studies suggest inhibition of ATPase activity of the CENP-E motor domain suppresses chromosome congression to the metaphase plate, subsequently SAC detects the misalignment of chromosomes and as a result, anaphase does not start.

Controlling CENP-E activity has a large potential to control mitosis progression. Although some potent CENP-E inhibitors have been developed so far, these inhibitors are not suitable for reversibly controlling CENP-E activity.

 

Fig.1 Overview of CENP-E in mitosis

Dr. Tamaoki, Dr. Uehara and co-workers, Hokkaido University, succeeded in obtaining the world’s first photoswitchable CENP-E inhibitor (PCEI-HU) which contains an azobenzene-based photo-isomerization unit. PCEI-HU can regulate CENP-E activity ON and OFF repeatedly by UV and visible light (Vis) irradiation.

 

PCEI-HU in water without light shows potent CENP-E inhibitory activity, but once UV light irradiation, PCEI-HU lacks its inhibitory activity. Under the condition, CENP-E keeps its motor activity for chromosome congression to the metaphase plate. This UV-induced loss of inhibitor action is recovered by Vis light. After Vis irradiation, PCEI-HU blocks CENP-E activity again and chromatin congression is clearly suppressed. PCEI-HU is a powerful tool for the fundamental investigation of prometaphase/metaphase and SAC.

 

Features 

・PCEI-HU works as CENP-E blocking form after visible light irradiation.

・PCEI-HU works as CENP-E non-blocking form after UV light irradiation.

・Until the all chromosomes are aligned to the metaphase plate in metaphase, movement can be regulated by visible / UV light.

 

Fig.2 Principle and overview of PCEI-HU

Data 

Fig.3 Absorption spectrum

Left: Absorption spectrum of PCEI-HU. Without light irradiation (w/o light; gray), after 365 nm light irradiation

        (UV irradiation; red) and after 510 nm light irradiation (Vis irradiation; blue).
Right : Absorbance changes at 338 nm under alternating 365/510 nm illumination.

 

Fig.4 Live cell imaging of mitotic chromosomes in PCEI-HU treated LLC-PK 1 cells

LLC-PK1 cells were treated with near-infrared DNA staining dye (SiR-DNA, 1 μM) and subsequently 1 μM PCEI-HU and 20 μM MG132 for 2 hours in darkroom. Cells were irradiated with UV light (365 nm) and Vis (510 nm) repeatedly and monitored in live-cell (Upper figure). 

Movement of a specific chromosome (yellow arrow) was analyzed in kymograph (Lower figure). While after UV irradiation, chromosomes moved to metaphase plate, but after Vis irradiation chromosomes left from metaphase plate. The chromosome was repeatedly regulated by UV/Vis cycles until chromosomes reaching to the metaphase plate.

 

* Research use only, not for human or animal therapeutic or diagnostic use !

 

References 

1. Mafy, N. N. et al., J. Am. Chem. Soc., 142, 1763-1767 (2020)

Photoswitchable CENP-E Inhibitor Enabling the Dynamic Control Chromosome Movement and Mitotic Progression.

 

코아사이언스 coresciences Funakoshi Inc. Japan 후나코시 한국 대리점 동원체 단백질-E 세포 분열 연구 염색체 이동 조절 단백질 탐색 억제제 억제자 염색체 이동 조절 시약