ABSTRACT
Fluorescent cell-permeant compounds based on 6-methoxy-8-p-toluenesulfonamido-quinoline, TSQ, are potentially powerful probes of intracellular zinc chemistry; however, the structure, thermodynamics, and stoichiometry of the metal complexes, and the molecular basis of Zn(II) recognition, remain open issues. To address these, we report the first structural characterization of a Zn(II) complex of a TSQ derivative, namely 2-methyl-6-methoxy-8-p-toluenesulfonamido-quinoline (3) and describe its unusual coordination chemistry. The crystal structure of the fluorescent complex of 3 with zinc reveals a 2:1 stoichiometry wherein bidentate coordination of two nitrogens from each ligand gives rise to a highly distorted tetrahedral Zn(II) center. Both sulfonamido groups in the zinc complex are tilted away from zinc to make room for coordination of the amide nitrogens. Zn-O(2) and Zn-O(4) distances are essentially nonbonding (3.06 and 3.10 A, respectively). The bond angles [N(1)-Zn-N(2) 83.5 degrees and N(3)-Zn-N(4) 83.0 degrees] are quite small relative to the 109 degrees angle of an ideal tetrahedral center. This result provides an insight into the zinc-binding mode of the TSQ derivative zinquin, in which a methyl group replaces the hydrogen in the 2-position of the quinoline ring. The methyl group and sulfonamide oxygen atoms clearly hinder formation of both square planar and octahedral complexes. We also show here that the Zn(II) complex of 3 in DMSO-water (80/20 w/w) exhibits an overall binding stability (log beta 2 = 18.24 +/- 0.02) similar to zinquin. Fluorescence microscopy suggests that each of these members of this family demarks a similar set of Zn(II)-enriched compartments that are common to all eukaryotic cells examined to date, and further shows that the ester function is not required for observation of these ubiquitous Zn-loaded compartments. The combined structural, thermodynamic, and physiological results provide a basis for design of other Zn(II)-specific membrane permeant probes with a range of Zn(II) affinities and photophysical properties.
Subject(s)
Aminoquinolines/chemistry , Aminoquinolines/metabolism , Tosyl Compounds/chemistry , Tosyl Compounds/metabolism , Zinc/metabolism , 3T3 Cells , Animals , Cell Line , Crystallography, X-Ray , Fluorescence , Magnetic Resonance Spectroscopy , Mass Spectrometry/methods , Mice , Mice, Inbred BALB C , Molecular Structure , Potentiometry , Zinc/chemistryABSTRACT
Reactive and potentially toxic cofactors such as copper ions are imported into eukaryotic cells and incorporated into target proteins by unknown mechanisms. Atx1, a prototypical copper chaperone protein from yeast, has now been shown to act as a soluble cytoplasmic copper(I) receptor that can adopt either a two- or three-coordinate metal center in the active site. Atx1 also associated directly with the Atx1-like cytosolic domains of Ccc2, a vesicular protein defined in genetic studies as a member of the copper-trafficking pathway. The unusual structure and dynamics of Atx1 suggest a copper exchange function for this protein and related domains in the Menkes and Wilson disease proteins.
Subject(s)
Carrier Proteins , Cation Transport Proteins , Copper/metabolism , Fungal Proteins/physiology , Molecular Chaperones/physiology , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/physiology , Amino Acid Sequence , Copper Transport Proteins , Escherichia coli , Fungal Proteins/metabolism , Humans , Molecular Sequence Data , Recombinant Proteins , Saccharomyces cerevisiae/metabolism , Sequence Homology, Amino AcidABSTRACT
The craniofacial characteristics and growth potential of 25 orthodontically treated patients with deep mandibular antegonial notch were compared with a similar group of 25 shallow notch subjects by the use of longitudinal lateral cephalometric radiographs. Deep notch cases had more retrusive mandibles with a shorter corpus, smaller ramus height, and a greater gonial angle than did shallow notch cases. The lower facial height in the subjects with a deep mandibular notch was found to be longer, and both the mandibular plane angle and facial axis were more vertically directed. During the average 4-year period examined, the deep notch sample experienced less mandibular growth as evidenced by a smaller increase in total mandibular length, corpus length, and less displacement of the chin in a horizontal direction than did the shallow notch sample. The results of this study suggest that the clinical presence of a deep mandibular antegonial notch is indicative of a diminished mandibular growth potential and a vertically directed mandibular growth pattern.
