Inducing rigid local structure around the zinc-binding region by hydrophobic interactions enhances the homotrimerization and apoptotic activity of zinc-free TRAIL

Lee, Hyun‐Wook; Kim, Tae-In; Chan, Khon Huynh; Kwon, Myung-Hee; Kim, Ji Soo; Jin, Moonsoo M.; Kim, Yong‐Sung

doi:10.1016/j.bbrc.2007.08.075

Cited by 8 publications

(10 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The construction of AY4 in a format of single-chain variable fragment, AY4-scFv, is described in Supplementary Materials and Methods. The experiments of size exclusion chromatography, ELISA, and surface plasmon resonance (SPR) of AY4 and AY4-scFv were done as described before (13,26,27). The detailed procedures are given in the figure legends and Supplementary Materials and Methods.…”

Section: Methodsmentioning

confidence: 99%

“…Z-VAD-fmk [benzyloxycarbonyl-Val-Ala-Asp-(OMe) fluoromethyl ketone] and Z-LEHD-fmk [Z-Leu-Glu(OMe)-His-Asp(OMe)-fmk] were from Santa Cruz Biotechnology. Soluble TRAIL (with a COOH-terminal 6× His-tag; described as TRAIL hereafter), DR5-ECD, DR4-ECD, DcR1-ECD, and DcR2-ECD were prepared as previously described (26,28). The ECD of CD95 was from Biovision.…”

Section: Methodsmentioning

confidence: 99%

“…29) was provided by Open Biosystems. The ECD (residues 53-169) of mTR was subcloned into pET21b (Novagen), expressed, and purified from Escherichia coli (26,28). Details for materials and protein preparations are provided in Supplementary Materials and Methods.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

A novel agonistic antibody to human death receptor 4 induces apoptotic cell death in various tumor cells without cytotoxicity in hepatocytes

Sung

Park²,

Lee³

et al. 2009

Molecular Cancer Therapeutics

Self Cite

View full text Add to dashboard Cite

The proapoptotic tumor necrosis factor-related apoptosis inducing ligand (TRAIL) receptors death receptor (DR) 4 and DR5 are attractive targets to develop the receptorspecific agonistic monoclonal antibodies (mAb) as anticancer agents because of their tumor-selective cell death-inducing activity. Here, we report a novel agonistic mAb, AY4, raised against human DR4 in mice. ELISA analysis revealed that AY4 specifically bound to DR4 without competition with TRAIL for the binding. Despite distinct binding regions of AY4 on DR4 from those of TRAIL, AY4 as a single agent induced caspase-dependent apoptotic cell death of several tumor types through the extrinsic and/or intrinsic pathways without substantial cytotoxicity to normal human hepatocytes. Further, the AY4-sensitive cells followed the same cell death characteristics classified as type I and type II cells by the response to TRAIL, suggesting that the cell death profiles in responses to DR4 and/or DR5 stimulation are determined by the downstream signaling of the receptor rather than the kind of receptor. Noticeably, AY4 efficiently induced cell death of Jurkat cells, which have been reported to be resistant to other anti-DR4 agonistic mAbs, most likely due to the unique epitope property of AY4. In vivo administration of AY4 significantly inhibited tumor growth of human non-small cell lung carcinoma preestablished in athymic nude mice. Conclusively, our results provide further insight into the DR4-mediated cell death signaling and potential use of AY4 mAb as an anticancer therapeutic agent, particularly for

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

A novel agonistic antibody to human death receptor 4 induces apoptotic cell death in various tumor cells without cytotoxicity in hepatocytes

Sung

Park²,

Lee³

et al. 2009

Molecular Cancer Therapeutics

Self Cite

View full text Add to dashboard Cite

show abstract

“…Protein concentrations were determined using the extinction coefficient of each protein, which were calculated from the amino acid sequence [8]. Subsequent biochemical analyses, such as SDS-PAGE, size-exclusion chromatography (SEC), and surface plasmon resonance (SPR), were also performed as essentially described previously [8,15] …”

Section: Modeling Of H3d8 Variable Fragment (Fv)mentioning

confidence: 99%

“…CD band intensities were expressed as mean residue ellipticity, [h], in mdeg cm 2 dmol À1 [16]. The unfolding experiments of m3D8 and h3D8 scFvs were measured using Trp fluorescence with a FP-6500 spectrofluorometer (Jasco Inc., Japan) after samples were equilibrated with various concentrations of GdnHCl (Sigma) overnight at 25°C [15]. The samples (10 lg/ ml in TBS) were excited at 295 nm, and the emission was monitored from 300 to 400 nm.…”

Section: (See Also Supplementary Figure Legends For Details)mentioning

confidence: 99%

Generation of humanized anti-DNA hydrolyzing catalytic antibodies by complementarity determining region grafting

Kim

Lee

Kim

et al. 2009

Biochemical and Biophysical Research Communications

Self Cite

View full text Add to dashboard Cite

A simplified method for the efficient purification and refolding of recombinant human TRAIL

Zhang

Hahn

Cao

et al. 2020

Biotechnology Progress

View full text Add to dashboard Cite

Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) belongs to the TNF cytokine superfamily that specifically induces apoptosis in a broad spectrum of human cancer cell lines but not in most healthy cells. The antitumor potential of recombinant human TRAIL (rhTRAIL) has attracted great attention among biologists and oncologists. However, attempts to express rhTRAIL in Escherichia coli often results in limited yield of bioactive protein due to the formation of inclusion bodies (IBs), which are dense insoluble particulate protein aggregates inside cells. We describe herein a highly simplified method to produce pure bioactive rhTRAIL using E. coli. The method is straightforward and requires only basic laboratory equipment, with highly efficient purification and high yield of renaturation, and may also be applied to produce other proteins that form IBs in E. coli.

show abstract

Inducing rigid local structure around the zinc-binding region by hydrophobic interactions enhances the homotrimerization and apoptotic activity of zinc-free TRAIL

Cited by 8 publications

References 15 publications

A novel agonistic antibody to human death receptor 4 induces apoptotic cell death in various tumor cells without cytotoxicity in hepatocytes

A novel agonistic antibody to human death receptor 4 induces apoptotic cell death in various tumor cells without cytotoxicity in hepatocytes

Generation of humanized anti-DNA hydrolyzing catalytic antibodies by complementarity determining region grafting

A simplified method for the efficient purification and refolding of recombinant human TRAIL

Contact Info

Product

Resources

About