Fluorescence Probes for Structure

Brand, Ludwig; Gohlke,

doi:10.1146/annurev.bi.41.070172.004211

Cited by 410 publications

(180 citation statements)

References 0 publications

Supporting

Mentioning

173

Contrasting

Order By: Relevance

“…The increase in TNS fluorescence is a well-established indicator for the exposure of hydrophobic regions on proteins (Brand and Gohlke, 1972) and has been observed previously with calpain (Hong et al, 1990). Thus, we argue that the exposure of a hydrophobic surface after calpain transition to its active conformation (Pal et al, 2001;Dainese et al, 2002;Li et al, 2004) should be responsible for the Ca 2q -dependent membrane binding in our experiments.…”

Section: Bereitgestellt Von | Universitaetsbibliothek Der Lmu Muenchensupporting

confidence: 81%

μ-Calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

Fernández-Montalván

Assfalg-Machleidt²,

Pfeiler³

et al. 2006

Biological Chemistry

View full text Add to dashboard Cite

active-site mutated heterodimeric human m-calpain with phospholipid bilayers was studied in vitro using proteinto-lipid fluorescence resonance energy transfer and surface plasmon resonance. Binding to liposomes was Ca 2q -dependent, but not selective for specific phospholipid head groups. wCa 2q x 0.5 for association with lipid bilayers was not lower than that required for the exposure of hydrophobic surface (detected by TNS fluorescence) or for enzyme activity in the absence of lipids. Deletion of domain V reduced the lipid affinity of the isolated small subunit (600-fold) and of the heterodimer (10-to 15-fold), thus confirming the proposed role of domain V for membrane binding. Unexpectedly, mutations in the acidic loop of the 'C2-like' domain III, a putative Ca 2q and phospholipid-binding site, did not affect lipid affinity. Taken together, these results support the hypothesis that in vitro membrane binding of m-calpain is due to the exposed hydrophobic surface of the active conformation and does not reduce the Ca 2q requirement for activation.

show abstract

Section: Bereitgestellt Von | Universitaetsbibliothek Der Lmu Muenchensupporting

confidence: 81%

μ-Calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

Fernández-Montalván

Assfalg-Machleidt²,

Pfeiler³

et al. 2006

Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…For these molecules the increase in fluorescence is assumed to originate from the rigid and/or hydrophobic environment at the binding site of the aggregates [30][31][32]. Although these compounds have been studied extensively, the fluorescence mechanism and the origin of the fluorescence increase upon binding are still controversial [31,[33][34][35].…”

Section: Absorption and Emission Propertiesmentioning

confidence: 99%

Anle138b and related compounds are aggregation specific fluorescence markers and reveal high affinity binding to α-synuclein aggregates

Deeg

Reiner

Schmidt

et al. 2015

Biochimica et Biophysica Acta (BBA) - General Subjects

View full text Add to dashboard Cite

Background: Special diphenyl-pyrazole compounds and in particular anle138b were found to reduce the progression of prion and Parkinson's disease in animal models. The therapeutic impact of these compounds was attributed to the modulation of α-synuclein and prion-protein aggregation related to these diseases. Methods: Photophysical and photochemical properties of the diphenyl-pyrazole compounds anle138b, anle186b and sery313b and their interaction with monomeric and aggregated α-synuclein were studied by fluorescence techniques. Results: The fluorescence emission of diphenyl-pyrazole is strongly increased upon incubation with α-synuclein fibrils, while no change in fluorescence emission is found when brought in contact with monomeric α-synuclein. This points to a distinct interaction between diphenyl-pyrazole and the fibrillar structure with a high binding affinity (K d = 190 ± 120 nM) for anle138b. Several α-synuclein proteins form a hydrophobic binding pocket for the diphenyl-pyrazole compound. A UV-induced dehalogenation reaction was observed for anle138b which is modulated by the hydrophobic environment of the fibrils. Conclusion: Fluorescence of the investigated diphenyl-pyrazole compounds strongly increases upon binding to fibrillar α-synuclein structures. Binding at high affinity occurs to hydrophobic pockets in the fibrils. General significance: The observed particular fluorescence properties of the diphenyl-pyrazole molecules open new possibilities for the investigation of the mode of action of these compounds in neurodegenerative diseases. The high binding affinity to aggregates and the strong increase in fluorescence upon binding make the compounds promising fluorescence markers for the analysis of aggregation-dependent epitopes.

show abstract

“…Sensitive physicochemical methods, such as fluorescence, have to be applied in order to obtain more structural and kinetic information. In other systems fluorescence techniques have been widely used because they offer high sensitivity and a wealth of information [6,7]. A pre- Cohn.…”

mentioning

confidence: 99%

Interactions of Yeast tRNA^Phe with Ribosomes from Yeast and Escherichia coli

Robertson

Kahan

Wintermeyer

et al. 1977

European Journal of Biochemistry

View full text Add to dashboard Cite

The interaction of ethidium-labeled tRNAPhe from yeast with ribosomes from yeast and Escherichiu coli was studied by steady-state measurements of fluorescence intensity and polarization. The ethidium label was covalently inserted into either the anticodon or the dihydrouridine loop of the tRNA. The codon-independent formation of a tRNA.ribosome complex led to only a moderate increase of the observed fluorescence polarization indicating a considerable internal mobility of the labeled parts of the tRNA molecule in the ribosome complex. When the ribosome complex was formed in the presence of poly(U), the probes both in the dihydrouridine loop and in the anticodon loop were strongly immobilized, the latter exhibiting a substantial increase in fluorescence intensity. A smaller intensity change was observed when E. coli ribosomes were used, although the extent of immobilization was found to be similar in this case. Competition experiments with non-labeled tRNAPhe showed that the labeled tRNA!& was readily released from the complex with yeast ribosomes when poly(U) was absent, whereas in the presence of poly(U) it was bound practically irreversibly. The finding that the mobility of a probe in the dihydrouridine loop is affected by the codon-anticodon interaction on the ribosome suggests a conformational change of the ribosome-bound tRNA which may involve opening of the tertiary structure interactions between the dihydrouridine and the TYC loop.In the past few years a rather detailed picture of the functional aspects of the tRNA-ribosome interaction has been elaborated [l -51. However, the physical basis of the high precision of protein synthesis and the mechanism of the individual steps have not yet been understood. Sensitive physicochemical methods, such as fluorescence, have to be applied in order to obtain more structural and kinetic information. In other systems fluorescence techniques have been widely used because they offer high sensitivity and a wealth of information [6,7]. A pre- Cohn. EtdBr, 2,7-diarnino-lO-ethyl-9-phenyl-phenanthridinium bromideorethidium bromide; t R N A h , a derivative of tRNAPhe in which wyebutine is replaced by ethidium; tRNA~~:Sl6;,,, tRNA"h' in which dihydrourdcil is replaced by ethidium; tRNAP_h;Wye, tRNAPhe lacking wyebutine.

show abstract

Fluorescence Probes for Structure

Cited by 410 publications

References 0 publications

μ-Calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

μ-Calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

Anle138b and related compounds are aggregation specific fluorescence markers and reveal high affinity binding to α-synuclein aggregates

Interactions of Yeast tRNA^Phe with Ribosomes from Yeast and Escherichia coli

Contact Info

Product

Resources

About

Fluorescence Probes for Structure

Cited by 410 publications

References 0 publications

μ-Calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

μ-Calpain binds to lipid bilayers via the exposed hydrophobic surface of its Ca2+-activated conformation

Anle138b and related compounds are aggregation specific fluorescence markers and reveal high affinity binding to α-synuclein aggregates

Interactions of Yeast tRNAPhe with Ribosomes from Yeast and Escherichia coli

Contact Info

Product

Resources

About

Interactions of Yeast tRNA^Phe with Ribosomes from Yeast and Escherichia coli