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M Martinez-Yamout

Publications and source records attributed to M Martinez-Yamout.

6 recordsLinked to original sources

Genomic-scale comparison of sequence- and structure-based methods of function prediction: does structure provide additional insight?

A function annotation method using the sequence-to-structure-to-function paradigm is applied to the identification of all disulfide oxidoreductases in the Saccharomyces cerevisiae genome. The method identifies 27 sequences as potential disulfide oxidoreductases. All previously known thioredoxins, glutaredoxins, and disulfide isomerases are correctly identified. Three of the 27 predictions are probable false-positives. Three novel predictions, which subsequently have been experimentally validated, are presented. Two additional novel predictions suggest a disulfide oxidoreductase regulatory mechanism for two subunits (OST3 and OST6) of the yeast oligosaccharyltransferase complex. Based on homology, this prediction can be extended to a potential tumor suppressor gene, N33, in humans, whose biochemical function was not previously known. Attempts to obtain a folded, active N33 construct to test the prediction were unsuccessful. The results show that structure prediction coupled with biochemically relevant structural motifs is a powerful method for the function annotation of genome sequences and can provide more detailed, robust predictions than function prediction methods that rely on sequence comparison alone.

Algorithms↗

Structure of the PHD zinc finger from human Williams-Beuren syndrome transcription factor.

The PHD (plant homeo domain) is a approximately 50-residue motif found mainly in proteins involved in eukaryotic transcription regulation. The characteristic sequence feature is a conserved Cys(4)-HisCys(3) zinc binding motif. We have determined the solution structure of the PHD motif from the human Williams-Beuren syndrome transcription factor (WSTF) protein. The domain folds into an interleaved zinc finger which binds two Zn(2+) in a similar manner to that of the RING and FYVE domains. The structure reveals a conserved zinc-binding core, together with two variable loops that are likely candidates for interactions between the various PHD domains and their specific ligands.

Amino Acid Sequence↗

Solution structure of the TAZ2 (CH3) domain of the transcriptional adaptor protein CBP.

The TAZ2 (CH3) domain of the transcriptional adapter protein CBP has been implicated in direct functional interactions with numerous cellular transcription factors and viral oncoproteins. The solution structure of the TAZ2 domain of murine CBP has been determined by nuclear magnetic resonance (NMR). The protein adopts a novel helical fold stabilized by three zinc ions, each of which is bound to one histidine and three cysteine ligands in HCCC-type motifs. Each zinc-binding site is formed from the carboxy terminus of an alpha-helix, a short loop, and the amino terminus of the next alpha-helix. A peptide derived from the N-terminal transactivation domain of p53 binds specifically to one face of the TAZ2 domain. The close similarities between the TAZ2 and TAZ1 (CH1 domain of CBP/p300) sequences suggest that both domains will adopt similar three-dimensional structures.

Adenovirus E1A Proteins↗

Solution structure of the cysteine-rich domain of the Escherichia coli chaperone protein DnaJ.

The solution structure of the cysteine-rich (CR) domain of Escherichia coli DnaJ has been solved by NMR methods. The structure of a 79 residue CR domain construct shows a novel fold with an overall V-shaped extended beta-hairpin topology. The CR domain is characterized by four C-X-X-C-X-G-X-G sequence motifs that bind two zinc ions. Residues in these two zinc modules show strong similarities in the grouping of resonances in the (15)N-(1)H HSQC spectrum and display pseudo-symmetry of the motifs in the calculated structures. The conformation of the cysteine residues coordinated to the zinc ion resembles that of the rubredoxin-knuckle, but there are significant differences in hydrogen bonding patterns in the two motifs. Zinc (15)N-(1)H HSQC titrations indicate that the fold of the isolated DnaJ CR domain is zinc-dependent and that one zinc module folds before the other. The C-X-X-C-X-G-X-G sequence motif is highly conserved in CR domains from a wide variety of species. The three-dimensional structure of the E. coli CR domain indicates that this sequence conservation is likely to result in a conserved structural motif.

Amino Acid Motifs↗

Site-directed mutagenesis and 1H nuclear magnetic resonance of an anti-dinitrophenyl spin label antibody.

Mutagenesis and spin label difference spectroscopy are used to assign the resonances of tyrosine residues in the binding site of the anti-dinitrophenyl spin label (DNP-SL) antibody AN02. Hapten binding constants of 13 point mutants are determined. From these studies it is clear that a light chain tyrosine specifically stabilizes DNP-SL binding, perhaps by means of a hydrogen bond to the hapten. This bond is absent in the case of the diamagnetic hapten dinitrophenyl-diglycine (DNP-Gly2). AN02 mutants with approximately 50 and 200-fold enhanced affinities for DNP-Gly2 are engineered. In these mutants, a single amino acid change relieves an electrostatic interaction between DNP-Gly2 and the binding site. The resulting improvement in hapten binding ability is specific to DNP-Gly2, since the affinity for DNP-SL is relatively unchanged. Evidence of conformational heterogeneity in the AN02/DNP-Gly2 complex is presented. In contrast with DNP-Gly, a ring proton of DNP-Gly2 experiences two environments on binding to AN02. It was previously shown that a light chain tyrosine (LY31) also assumes two conformations when DNP-Gly2 is bound. The simultaneous presence of multiple AN02/DNP-Gly2 complexes implies conformational isomerism of a tryptophan residue which contacts both the hapten and LY31.

Amino Acid Sequence↗