Amphidinolide H, a Potent Cytotoxic Macrolide, Covalently Binds on Actin Subdomain 4 and Stabilizes Actin Filament

Usui, Takeo; Kazami, Sayaka; Dohmae, Naoshi; Mashimo, Yoshikazu; Kondo, Hideki; Tsuda, Masashi; Terasaki, Asako G.; Ohashi, Kazuyo; Kobayashi, Jun’ichi; Osada, Hiroyuki

doi:10.1016/j.chembiol.2004.07.014

Cited by 77 publications

(78 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MALDI-TOF mass analysis and the halo assay using the yeast harboring site-directed mutagenized actin revealed that the covalent binding of amphidinolide H (8) of small actin patches, followed by F-actin rearrangement into aggregates via the retraction actin fibers. These results indicated that amphidinolide H (8) is a novel F-actin stabilizer that covalently binds on actin [45].…”

Section: Mechanism Of Action Of Amphidinolides B and Hmentioning

confidence: 83%

“…These results suggest that amphidinolide B (2) enhances an interaction of actin and myosin directly and increases Ca 2ϩ sensitivity of the contractile apparatus mediated through troponin-tropomyosin system, resulting in an increase in the ATPase activity of actomyosin and thus enhances the contractile response of myofilament [44]. The molecular target of amphidinolide H (8) [25,26] has been investigated as follows [45,46]. The analysis of phenotypes of amphidinolide H-treated cells suggested that amphidinolide H (8) disrupt the actin organization in the cells, and the polymerization/depolymerization assay using purified actin indicated that amphidinolide H (8) stimulated actin polymerization and stabilized F-actin.…”

Section: Mechanism Of Action Of Amphidinolides B and Hmentioning

confidence: 99%

“…Recently, eleven new macrolides, amphidinolides B4 (35), B5 (36), and C2 (37), amphidinolactones A (38) and B (39), iriomoteolides 1aϳ1c (40ϳ42) and 3a (43), and amphidinolides B6 (45) and B7 (44), (Table 1) have been isolated from five strains (Y-100, Y-71, Y-25, HYA024, and HYA002) of the genus Amphidinium, which were collected at different location in Okinawa. This review covers recent progress on the isolation and structure elucidation of these cytotoxic macrolides as well as the molecular target of amphidinolide H (8), cloning of the putative polyketide synthase (PKS) gene of amphidinolide biosynthesis, and synthesis of some amphidinolides recently achieved by other groups.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Amphidinolides and Its Related Macrolides from Marine Dinoflagellates

Kobayashi

2008

J Antibiot

Self Cite

145

View full text Add to dashboard Cite

This review covers the recent results described in our publications on several new cytotoxic macrolides isolated from dinoflagellates of the genus Amphidinium in addition to an overview of the isolation, structure elucidation, synthesis, biosynthesis, and bioactivity of a series of cytotoxic macrolides, named amphidinolides, reported so far. IntroductionMarine dinoflagellates, a diverse group of unicellular eukaryotes, have been recognized as real producers of marine toxins responsible to fish and algal poisoning, as well as biologically unique organisms due to their taxonomic position and unusual chromosome structure and composition. Marine dinoflagellates have also proved to be one of the most important source of bioactive natural products, which have been investigated worldwide. We have continuously studied structurally intriguing and biologically interesting bioactive macrolides and polyketides from dinoflagellates Amphidinium sp., which are symbionts of Okinawan marine flatworms Amphiscolops sp.Thirty Culture of Dinoflagellates Amphidinium sp.Large-scale cultures of the dinoflagellates of the genus Amphidinium have been performed using seawater medium enriched with Provasoli's Erd-Schriber (ES) supplement. Static incubation with illumination in a cycle of 16 hours of light and 8 hours of darkness was carried out for two weeks at 25°C. The cultures were harvested by removal of the supernatant through suction and then centrifugation to obtain algal cells. Harvested cells were extracted with MeOH -toluene, and the extracts were subjected to a systematic separation using several chromatographies to yield cytotoxic macrolides [15ϳ26]. Isolation of New Macrolides 3-1. Amphidinolactones A and BA new 13-membered macrolide, amphidinolactone A (38), C 20 H 30 O 4 , was isolated from a strain (Y-25) of a dinoflagellate Amphidinium sp. [15]. Detailed analyses of the 1 H-1 H COSY spectrum of 38 revealed connectivities of a long carbon chain from C-2 to C-20. 1 H and 13 C chemical shifts of CH 2 -2 and CH-12 suggested that C-12 was involved in an ester linkage with C-1. 1 H-1 H couplings of the two disubstituded double bonds at C-5 and C-9 indicated the Z and E geometries, respectively. Geometries of two disubstituted double bonds at C-14 and C-17 were assigned as both Z by NOESY correlations and the carbon chemical shift of C-16, which was a typical value for a methylene carbon between two Z double bonds. Thus, the gross structure of amphidinolactone A was elucidated to be 38. The relative configurations at C-8, C-11, and C-12 in 38 were deduced from NOESY correlations.A new 26-membered macrolide, amphidinolactone B (39), C 32 H 54 O 8 , has been isolated from the same dinoflagellate Amphidinium sp. (strain Y-25) as described above [16]. Detailed analyses of the 1 H-1 H COSY and TOCSY spectra as well as HMBC correlations indicated connectivities of a long carbon chain from C-2 to C-26. 1 H and 13 C chemical shifts of C-25 indicated that C-25 was involved in an ester linkage with C-1. The NOESY correlation for...

show abstract

Section: Mechanism Of Action Of Amphidinolides B and Hmentioning

confidence: 83%

Section: Mechanism Of Action Of Amphidinolides B and Hmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Amphidinolides and Its Related Macrolides from Marine Dinoflagellates

Kobayashi

2008

J Antibiot

Self Cite

145

View full text Add to dashboard Cite

show abstract

“…In the cultured mammalian cells, both cause actin filament aggregation (Hoehn et al 1973;Usui et al 2004). In this study, we report the purification of Cuc E from A. macrocarpa and evaluated direct interaction between Cuc E and purified actin.…”

Section: Introductionmentioning

confidence: 99%

Direct interaction of Cucurbitacin E isolated from Alsomitra macrocarpa to actin filament

et al. 2007

View full text Add to dashboard Cite

A methanol extract of Alsomitra macrocarpa leaves and branches induced a marked alteration of cell morphology in a human stellate cell line (LX-2). Similar morphologic alterations were observed in several other cell lines. Active compound was purified from the extract and determined to be cucurbitacin E (Cuc E). It has been known that Cuc E causes marked disruption of the actin cytoskeleton, supporting our observation, but how Cuc E altered the actin cytoskeleton has not been elucidated. By using the standard fluorescence assay using copolymerization and depolymerization of native and pyrene labelled actin, this study revealed that Cuc E interacted directly with actin consequently stabilizing the polymerized actin. When NIH-3T3 cells exogenously expressing YFP-labeled actin were treated with Cuc E, firstly the aggregation of globular actin and secondly the aggregation of actin including disrupted fibrous actin in the cells was observed.

show abstract

“…Interestingly it not only enhances phalloidin-binding to F-actin, but also binds to both F-and G-actins with similar affinities of 0.21 μM and 0.43 μM, respectively (Saito et al 2004). Amphidinolide H covalently binds to Tyr-198 of the actin monomer (Usui et al 2004). Tyr-198 is in subdomain IV, which faces toward the phalloidin-binding gap (Oda et al 2005).…”

Section: Marine Toxins That Stabilize Actin Filamentsmentioning

confidence: 99%

Developmental Biology of Neoplastic Growth

Macieira‐Coelho¹

2005

Progress in Molecular and Subcellular Biology

View full text Add to dashboard Cite

Amphidinolide H, a Potent Cytotoxic Macrolide, Covalently Binds on Actin Subdomain 4 and Stabilizes Actin Filament

Cited by 77 publications

References 46 publications

Amphidinolides and Its Related Macrolides from Marine Dinoflagellates

Amphidinolides and Its Related Macrolides from Marine Dinoflagellates

Direct interaction of Cucurbitacin E isolated from Alsomitra macrocarpa to actin filament

Developmental Biology of Neoplastic Growth

Contact Info

Product

Resources

About