Metal Compounds as Tools for the Construction and the Interpretation of Medium-Resolution Maps of Ribosomal Particles

Weinstein, S.; Jahn, Werner; Glotz, Carola; Schlünzen, Frank; Levin, Inna; Janell, Daniela; Harms, Joerg; Kölln, Ingo; Hansen, Harly A. S.; Glühmann, Marco; Bennett, William S.; Bartels, Heike; Bashan, Anat; Agmon, Ilana; Kessler, Maggie; Pioletti, Marta; Avila, Horacio; Αναγνωστόπουλος, Κωνσταντίνος; Peretz, Moshe; Auerbach, Tamar; Franceschi, François; Yonath, Ada

doi:10.1006/jsbi.1999.4135

Cited by 29 publications

(34 citation statements)

References 54 publications

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“…This small cluster may be useful for higher-resolution labeling and as an isomorphous replacement in the X-ray phasing of large unit cell protein crystals (Weinstein et al 1989;Weinstein et al 1999). …”

Section: Tetra Iridiummentioning

confidence: 99%

New Frontiers in Gold Labeling

Hainfeld

Powell

2000

J Histochem Cytochem.

239

173

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Recent advances in gold technology have led to probes with improved properties and performance for cell biologists: higher labeling density, better sensitivity, and greater penetration into tissues. Gold clusters, such as the 1.4-nm Nanogold, are gold compounds that can be covalently linked to Fab′ antibody fragments, making small and stable probes. Silver enhancement then makes these small gold particles easily visible by EM, LM, and directly by eye. Another advance is the combination of fluorescent and gold probes for correlative microscopy. Chemical crosslinking of gold particles to many biologically active molecules has made possible many novel probes, such as gold-lipids, gold-Ni-NTA, and gold-ATP.

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Section: Tetra Iridiummentioning

confidence: 99%

New Frontiers in Gold Labeling

Hainfeld

Powell

2000

J Histochem Cytochem.

239

173

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confidence: 99%

“…We have commercialized combined fluorescent and heavy metal probes that enable collection of complimentary sets of data, from fluorescence and electron microscopy, for correlative microscopic studies of biological targets at different spatial resolutions [3]. We herein report combined heavy metal probes that will enable detection and spatially resolved chemical analysis of biological samples.Recently, we and others reported construction of electron density maps of tetra-iridium (Ir 4 ) cluster labeled virus capsids and ribosome particles by cryo-EM and X-ray crystallography respectively at medium to high resolution [1, 4]. These Ir 4 clusters contain carbonyl ligands that exhibit extremely intense infrared (IR) absorptions in the region where most of the biological materials do not absorb (Fig.…”

mentioning

confidence: 99%

“…Heavy metal cluster complex labels that can be derivatized for sitespecific covalent attachment to macromolecules have clear advantages over heavy metal salts [1,2]. We have commercialized combined fluorescent and heavy metal probes that enable collection of complimentary sets of data, from fluorescence and electron microscopy, for correlative microscopic studies of biological targets at different spatial resolutions [3].…”

mentioning

confidence: 99%

“…Recently, we and others reported construction of electron density maps of tetra-iridium (Ir 4 ) cluster labeled virus capsids and ribosome particles by cryo-EM and X-ray crystallography respectively at medium to high resolution [1, 4]. These Ir 4 clusters contain carbonyl ligands that exhibit extremely intense infrared (IR) absorptions in the region where most of the biological materials do not absorb (Fig.…”

mentioning

confidence: 99%

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High Z Metal Carbonyls for Imaging and Microspectroscopy

Joshi

Ramamurthy

Powell

et al. 2005

MAM

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The power of microscopic and crystallographic techniques in structure-function analysis of multi-protein complexes can be greatly enhanced by the use of high Z or heavy atom labels introduced at specific sites [1]. Heavy metal cluster complex labels that can be derivatized for sitespecific covalent attachment to macromolecules have clear advantages over heavy metal salts [1,2]. We have commercialized combined fluorescent and heavy metal probes that enable collection of complimentary sets of data, from fluorescence and electron microscopy, for correlative microscopic studies of biological targets at different spatial resolutions [3]. We herein report combined heavy metal probes that will enable detection and spatially resolved chemical analysis of biological samples.Recently, we and others reported construction of electron density maps of tetra-iridium (Ir 4 ) cluster labeled virus capsids and ribosome particles by cryo-EM and X-ray crystallography respectively at medium to high resolution [1, 4]. These Ir 4 clusters contain carbonyl ligands that exhibit extremely intense infrared (IR) absorptions in the region where most of the biological materials do not absorb (Fig. 1b). This unique feature of metal carbonyls has led to the development of non-isotopic IR spectroscopic immunoassays [5]. We recently demonstrated that functionalized heavy atom carbonyl cluster labeled reagents can be used for bio-sensing/bio-recognition using Fourier transform infrared (FTIR) spectroscopy, albeit with lower detection sensitivities than competing technologies [6]. To demonstrate the use of heavy metal carbonyl clusters as dual labels for electron microscopy and FTIR microspectroscopy, we conjugated the previously reported Ir 4 label [4] to freshly reduced goat anti-rabbit F(ab') fragments following standard protocols [7]. The Ir 4 labels are clearly visible in dark-field scanning transmission electron micrograph (STEM) of the Ir 4 -labeled F(ab') conjugate (Fig. 2a). The Ir 4 -F(ab') conjugate was captured by 100 ng of covalently immobilized rabbit IgG on IR reflective slides following reported procedures [6,8]. The Ir 4 labels could be localized at 10x10 micron resolution by FTIR microspectroscopy (Fig. 2b).Since IR microspectroscopic imaging has been used to distinguish chemical differences between healthy and diseased tissues [9], we envisage the use of metal carbonyl cluster labels for rapid localization of diseased tissues. Subtraction of the Ir-4 label spectrum from that of localized targets will decipher chemical information and underlying chemistry which other methods, such as light or fluorescence microscopy, do not. The Ir 4 labels reported here have active Raman vibrations. Since Raman microspectroscopy has also been used to distinguishing healthy and diseased tissues, we are investigating the use of metal carbonyls as labels for Raman microspectroscopy.

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Cysteine‐Specific, Covalent Anchoring of Transition Organometallic Complexes to the Protein Papain from Carica papaya

et al. 2007

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Site-directed and covalent introduction of various transition metal-organic entities to the active site of the cysteine endoproteinase, papain, was achieved by treatment of this enzyme with a series of organometallic maleimide derivatives specially designed for the purpose. Kinetic studies made it clear that time-dependent irreversible inactivation of papain occurred in the presence of these organometallic maleimides as a result of Michael addition of the sulfhydryl of Cys25. The rate and mechanism of inactivation were highly dependent on the structure of the organometallic entity attached to the maleimide group. Combined ESI-MS and IR analysis indicated that all the resulting papain adducts contained one organometallic moiety per protein molecule. This confirmed that chemospecific introduction of the metal complexes was indeed achieved. Thus, three novel reagents for heavy-atom derivatization of protein crystals, which include ruthenium, rhenium and tungsten, are now available for the introduction of electron-dense scatterers for phasing of X-ray crystallographic data.

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Metal Compounds as Tools for the Construction and the Interpretation of Medium-Resolution Maps of Ribosomal Particles

Cited by 29 publications

References 54 publications

New Frontiers in Gold Labeling

New Frontiers in Gold Labeling

High Z Metal Carbonyls for Imaging and Microspectroscopy

Cysteine‐Specific, Covalent Anchoring of Transition Organometallic Complexes to the Protein Papain from Carica papaya

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