ChemInform Abstract: Polypeptide ‐ Metal Cluster Connectivities in Metallothionein 2 by Novel <sup>1</sup>H ‐ <sup>113</sup>Cd Heteronuclear Two‐Dimensional NMR Experiments

Frey, M. H.; Wagner, Gerhard; Vašák, Milan; Soerensen, O. W.; Neuhaus, David; Woergoetter, Erich.; Kaegi, Jeremias H. R.; Ernst, R. R.; Wuethrich, Kurt

doi:10.1002/chin.198612051

Cited by 2 publications

(2 citation statements)

References 1 publication

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since active sites and other binding sites usually are located on the surface of the proteins, very important regions of the protein may be distorted. Some structures even show large differences between NMR and X-ray structure (Frey et al, 1985;Klevit and Waygood, 1986) The advantage of NMR is that it is dealing with protein molecules in solution, usually in an environment not too different from its natural one. It is possible to study the protein and the dynamical aspects of its interaction with other molecules like substrates, inhibitors, etc.…”

Section: Nmr Of Proteinsmentioning

confidence: 99%

The blind watchmaker and rational protein engineering

Anthonsen

Baptista

Drabløs

et al. 1994

Journal of Biotechnology

View full text Add to dashboard Cite

In the present review some scientific areas of key importance for protein engineering are discussed, such as problems involved in deducting protein sequence from DNA sequence (due to posttranscriptional editing, splicing and posttranslational modifications), modelling of protein structures by homology, NMR of large proteins (including probing the molecular surface with relaxation agents), simulation of protein structures by molecular dynamics and simulation of electrostatic effects in proteins (including pH-dependent effects). It is argued that all of these areas could be of key importance in most protein engineering projects, because they give access to increased and often unique information. In the last part of the review some potential areas for future applications of protein engineering approaches are discussed, such as non-conventional media, de novo design and nanotechnology.

show abstract

Section: Nmr Of Proteinsmentioning

confidence: 99%

The blind watchmaker and rational protein engineering

Anthonsen

Baptista

Drabløs

et al. 1994

Journal of Biotechnology

View full text Add to dashboard Cite

show abstract

“…Mammalian MTs contain 60–68 amino acids with an absolutely conserved pattern of 20 cysteines [2]. The three‐dimensional structure of mammalian MT has been determined for MT metalloforms with seven divalent metal ions (Zn 2+ and/or Cd 2+ ) by NMR [3–5] and by X‐ray diffraction [6]. The NMR and the crystal structures are similar and they expose two protein domains, both folded into metal‐thiolate clusters [7].…”

mentioning

confidence: 99%

The effects of physiologically important nonmetallic ligands in the reactivity of metallothionein towards 5,5′‐dithiobis(2‐nitrobenzoic acid)

Kangur

Palumaa²

2001

European Journal of Biochemistry

View full text Add to dashboard Cite

The reaction of Cd 5 Zn 2 -metallothionein (MT) with 5,5 0 -dithiobis(2-nitrobenzoic acid) (Nbs 2 ) has been studied at different reagent stoichiometries, pH and temperature conditions and in the presence of several ligands. At stoichiometries of Nbs 2 to MT from 0.5 to 5, the reaction followed first order kinetics. The first order rate constants obtained were independent from the concentration of Nbs 2 but were linearly dependent on the concentration of MT. At higher Nbs 2 /MT stoichiometries, the reaction deviates from first order kinetics and the observed rate constant increases. The reactivity of MT towards Nbs 2 has been probed at 4 mM concentration of both reagents where the reaction is monophasic and is characterized by a linear Arrhenius plot (E a ¼ 45.8^2.7 kJ : mol .07 mM. Several compounds such as AMP, S-methylglutathione, and phosphate had no effect on the reaction, but Zn 21 ions showed an inhibitory effect at micromolar concentrations.Keywords: metallothionein; Ellman's reagent; 5,5 0 -dithiobis(2-nitrobenzoic acid); ATP; reactivity of metal thiolate clusters.Metallothioneins (MTs) represent a class of small cysteinerich proteins adapted for the binding of various 'soft' transition metal ions into metal (thiolate clusters) [1]. Mammalian MTs contain 60 -68 amino acids with an absolutely conserved pattern of 20 cysteines [2]. The threedimensional structure of mammalian MT has been determined for MT metalloforms with seven divalent metal ions (Zn 21 and/or Cd 21 ) by NMR [3 -5] and by X-ray diffraction [6]. The NMR and the crystal structures are similar and they expose two protein domains, both folded into metal-thiolate clusters [7]. The N-terminal b domain (amino acids 1 -31) contains a cluster of three metals co-ordinated with nine thiolates, and the C-terminal a domain contains a cluster of four metals co-ordinated with 11 thiolates [7,8].MT is a ubiquitous protein and in cells it is induced by a variety of stimuli, including metals, hormones, chemical and physical stress factors [1,9]. However, the primary physiological role of MT is unclear and is still a matter for discussion [10,11]. At the same time, it is generally accepted that MTs are multifunctional proteins participating in the metabolism of zinc and copper, in detoxification of toxic metals such as cadmium and mercury, and in the protection of cells against reactive oxygen species and other electrophiles [9,12]. Some tissue-specific MT isoforms may also have additional physiological functions, such as inhibition of neuronal growth in the case of brain-specific 14].Metal transfer and release play an important role in functioning of MT and the appropriate mechanisms have been intensively studied. Generally, MT-bound metals are kinetically labile [15] and could be transferred to other proteins or released into the environment. It has been demonstrated that MT can participate in metal transfer and/or exchange with different metalloproteins including enzymes [16][17][18], receptors [19] and transcription factors [20]. The mechanisms ...

show abstract

ChemInform Abstract: Polypeptide ‐ Metal Cluster Connectivities in Metallothionein 2 by Novel ¹H ‐ ¹¹³Cd Heteronuclear Two‐Dimensional NMR Experiments

Cited by 2 publications

References 1 publication

The blind watchmaker and rational protein engineering

The blind watchmaker and rational protein engineering

The effects of physiologically important nonmetallic ligands in the reactivity of metallothionein towards 5,5′‐dithiobis(2‐nitrobenzoic acid)

Contact Info

Product

Resources

About

ChemInform Abstract: Polypeptide ‐ Metal Cluster Connectivities in Metallothionein 2 by Novel 1H ‐ 113Cd Heteronuclear Two‐Dimensional NMR Experiments

Cited by 2 publications

References 1 publication

The blind watchmaker and rational protein engineering

The blind watchmaker and rational protein engineering

The effects of physiologically important nonmetallic ligands in the reactivity of metallothionein towards 5,5′‐dithiobis(2‐nitrobenzoic acid)

Contact Info

Product

Resources

About

ChemInform Abstract: Polypeptide ‐ Metal Cluster Connectivities in Metallothionein 2 by Novel ¹H ‐ ¹¹³Cd Heteronuclear Two‐Dimensional NMR Experiments