High metal substitution tolerance of anthrax lethal factor and characterization of its active copper-substituted analogue

Lo, Suet Y.; Säbel, Crystal E.; Webb, Michael I.; Walsby, Charles J.; Siemann, Stefan

doi:10.1016/j.jinorgbio.2014.06.009

Cited by 10 publications

(12 citation statements)

References 80 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…On the other hand, the donation of Cd(II) often causes Zn-metalloenzymes to partially or completely lose function. [63][64][65] By locking most of the Cd(II) in a cluster structure, donation may be discouraged. Only a small change in pH would induce clustering and many biological compartments have a pH well below 7.2 where clustering begins to take over as the preferred Cd-binding pathway.…”

Section: Understanding Mt1a's Multitude Of Biological Functionsmentioning

confidence: 99%

Defining the metal binding pathways of human metallothionein 1a: balancing zinc availability and cadmium seclusion

2016

View full text Add to dashboard Cite

Metallothioneins (MTs) are cysteine-rich, metal-binding proteins that are found throughout Nature. This ubiquity highlights their importance in essential metal regulation, heavy metal detoxification and cellular redox chemistry. Missing from the current description of MT function is the underlying mechanism by which MTs achieve their proposed biological functions. To date, there have been conflicting reports on the mechanism of metal binding and the structures of the metal binding intermediates formed during metalation of apoMTs. The form of the metal-bound intermediates dictates the metal sequestering and metal-donating properties of the protein. Through a detailed analysis of spectral data from electrospray ionization mass spectromeric and circular dichroism methods we report that Zn(ii) and Cd(ii) metalation of the human MT1a takes place through two distinct pathways. The first pathway involves formation of beaded structures with up to five metals bound terminally to the 20 cysteines of the protein via a noncooperative mechanism. The second pathway is dominated by the formation of the four-metal domain cluster structure M4SCYS11via a cooperative mechanism. We report that there are different pathway preferences for Zn(ii) and Cd(ii) metalation of apo-hMT1a. Cd(ii) binding follows the beaded pathway above pH 7.1 but beginning below pH 7.1 the clustered (Cd4Scys11) pathway begins to dominate. In contrast, Zn(ii) binding follows the terminal, "beaded", pathway at all physiologically relevant pH (pH ≥ 5.2) only following the clustered pathway below pH 5.1. The results presented here allow us to reconcile the conflicting reports concerning the presence of different metalation intermediates of MTs. The conflict regarding cooperative versus noncooperative binding mechanisms is also reconciled with the experimental results described here. These two metal-specific pathways and the presence of radically different intermediate structures provide insight into the multi-functional nature of MT: binding Zn(ii) terminally for donation to metalloenzymes and sequestering toxic Cd(ii) in a cluster structure.

show abstract

Section: Understanding Mt1a's Multitude Of Biological Functionsmentioning

confidence: 99%

Defining the metal binding pathways of human metallothionein 1a: balancing zinc availability and cadmium seclusion

2016

View full text Add to dashboard Cite

show abstract

“…In light of these considerations, the current study sought to probe the effect of arginine-containing peptides on the function of VIM-2, a member of the clinically prevalent VIM-group of MBLs [51][52][53]. The impetus for this investigation stemmed from our initial observation that a peptide which harbours an octaarginine core and serves as a prototypical substrate of the unrelated zinc protease anthrax lethal factor [31,54] was capable of inhibiting VIM-2 at sub-micromolar concentrations. Hence, the current study was aimed at elucidating the structure-function relationship between this enzyme and a variety of arginine-bearing peptides.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, the inhibition by these peptides is dependent not only on the substrate but also on the nature of the MBL, with IMP-1 being much less sensitive to inhibition than VIM-2. Furthermore, the original lead compound (peptide 1) is a well-studied substrate of the anthrax lethal factor rather than an inhibitor of this zinc-dependent enzyme [31,54].…”

Section: Accepted Manuscriptmentioning

confidence: 99%

Arginine-containing peptides as potent inhibitors of VIM-2 metallo-β-lactamase

Rotondo

Marrone

Goodfellow

et al. 2015

Biochimica et Biophysica Acta (BBA) - General Subjects

Self Cite

View full text Add to dashboard Cite

“…1 Demetallated LF (apoLF) was prepared by treatment of ZnLF with ethylenediaminetetraacetic acid (EDTA) and dipicolinic acid (DPA) as documented previously. 2 Cobalt-substituted LF (CoLF) was obtained using either the direct exchange method 1 or by reconstitution of apoLF with CoCl2 according to the protocols outlined previously. 2 Copper-substituted LF (CuLF) was prepared by apoLF reconstitution with CuSO4 as reported in the literature.…”

Section: Preparation Of Lfmentioning

confidence: 99%

Highly dynamic metal exchange in anthrax lethal factor involves the occupation of an inhibitory metal binding site

Young

Siemann

2016

Chem. Commun.

Self Cite

View full text Add to dashboard Cite

show abstract

High metal substitution tolerance of anthrax lethal factor and characterization of its active copper-substituted analogue

Cited by 10 publications

References 80 publications

Defining the metal binding pathways of human metallothionein 1a: balancing zinc availability and cadmium seclusion

Defining the metal binding pathways of human metallothionein 1a: balancing zinc availability and cadmium seclusion

Arginine-containing peptides as potent inhibitors of VIM-2 metallo-β-lactamase

Highly dynamic metal exchange in anthrax lethal factor involves the occupation of an inhibitory metal binding site

Contact Info

Product

Resources

About