113Cd NMR Experiments Reveal an Unusual Metal Cluster in the Solution Structure of the Yeast Splicing Protein Bud31p†
نویسندگان
چکیده
Establishing the binding topology of structural zinc ions in proteins is an essential part of their structure determination by NMR spectroscopy. Using 113Cd NMR experiments with 113Cd-substituted samples is a useful approach but has previously been limited mainly to very small protein domains. Here we used 113Cd NMR spectroscopy during structure determination of Bud31p, a 157-residue yeast protein containing an unusual Zn3Cys9 cluster, demonstrating that recent hardware developments make this approach feasible for significantly larger systems.
منابع مشابه
NMR analysis of the structure and metal sequestering properties of metallothioneins.
113Cd-NMR studies have been used to elucidate the structure of the metal-binding sites in mammalian and invertebrate ( Scylla serrata) metallothioneins (MTs). Chemical shift data have shown that all Cd ions are tetrahedrally coordinated to four cysteine thiolate ligands with single cysteinyl sulfurs bridging adjacent metals. Homonuclear decoupling experiments have shown that the 7 g-atoms of me...
متن کاملDynamic metal-thiolate cluster structure of metallothioneins.
113Cd-NMR studies have established that vertebrate and invertebrate metallothioneins contain two unique metal-thiolate clusters. However, it proved to be difficult to account theoretically for all features of the 113Cd-NMR spectra. In a reinvestigation of these features using chromatographically homogeneous 113Cd7-metallothionein we have identified a total of seven 113Cd resonances and have con...
متن کاملGAL4 transcription factor is not a "zinc finger" but forms a Zn(II)2Cys6 binuclear cluster.
The DNA-binding domain of the transcription factor GAL4, consisting of the 62 N-terminal residues and denoted GAL4(62*), contains a Cys-Xaa2-Cys-Xaa6-Cys-Xaa6-Cys-Xaa2-Cys-Xaa6+ ++-Cys motif, which has been shown previously to bind two Zn(II) or Cd(II) ions. Binding of Zn(II) or Cd(II) is essential for the recognition by GAL4 of the specific palindromic DNA sequence to which it binds upstream o...
متن کاملNMR studies of crab and plaice metallothioneins.
Metallothioneins isolated from the hepatopancreas of the edible crab (Cancer paqurus) and the plaice (Pleuronectes platessa) after cadmium injection are predominantly cadmium proteins containing only small amounts of zinc and traces of copper. The removal of metal ions from the two metallothioneins by EDTA was studied using proton NMR spectroscopy. The rates of removal of cadmium and zinc were ...
متن کاملInteractions of the yeast SF3b splicing factor.
The U2 snRNP promotes prespliceosome assembly through interactions that minimally involve the branchpoint binding protein, Mud2p, and the pre-mRNA. We previously showed that seven proteins copurify with the yeast (Saccharomyces cerevisiae) SF3b U2 subcomplex that associates with the pre-mRNA branchpoint region: Rse1p, Hsh155p, Hsh49p, Cus1p, and Rds3p and unidentified subunits p10 and p17. Here...
متن کامل