Biomedical subjects

Christopher C Fischer

Publications and source records attributed to Christopher C Fischer.

3 recordsLinked to original sources

PubMed2006

Predicting crystal structure by merging data mining with quantum mechanics.

Modern methods of quantum mechanics have proved to be effective tools to understand and even predict materials properties. An essential element of the materials design process, relevant to both new materials and the optimization of existing ones, is knowing which crystal structures will form in an alloy system. Crystal structure can only be predicted effectively with quantum mechanics if an algorithm to direct the search through the large space of possible structures is found. We present a new approach to the prediction of structure that rigorously mines correlations embodied within experimental data and uses them to direct quantum mechanical techniques efficiently towards the stable crystal structure of materials.

Journal Article↗

PubMed2004

Palmitoylation and plasma membrane targeting of RGS7 are promoted by alpha o.

Regulator of G protein signaling (RGS) proteins modulate G protein signaling by acting as GTPase-activating proteins for G protein alpha-subunits. RGS7 belongs to a subfamily of RGS proteins that exist as dimers with the G protein beta(5)-subunit. In this report, we addressed the mechanisms of plasma membrane localization of beta(5)RGS7. When expressed in human embryonic kidney 293 cells, beta(5)RGS7 was found to be cytoplasmic and soluble. Expression of alpha(o) promoted a strong redistribution of beta(5)RGS7 to the plasma membrane. Expression of alpha(q), however, failed to affect the subcellular localization of beta(5)RGS7. The constitutively active mutant alpha(o)R179C, like wild-type alpha(o), strongly recruited beta(5)RGS7 to plasma membranes; however, inactive alpha(o)G204A, RGS-insensitive alpha(o)G184S, and lipidation-deficient alpha(o)G2A were all defective in the ability to promote plasma membrane localization of beta(5)RGS7. In addition, palmitoylation of RGS7 was demonstrated, and palmitoylation required expression of alpha(o) or alpha(o)R179C. To examine potential palmitoylation sites of RGS7, several cysteines were substituted with serines. beta(5)RGS7C133S failed to localize to plasma membranes when coexpressed with alpha(o), suggesting cysteine 133 of RGS7 as a putative palmitoylation site. Finally, deletion of amino acids 76 to 128 of RGS7, which includes part of the disheveled, EGL-10, pleckstrin (DEP) domain, prevented alpha(o)-mediated plasma membrane recruitment of beta(5)RGS7. These findings are the first to demonstrate Galpha-regulated plasma membrane localization and palmitoylation of beta(5)RGS7 and suggest that membrane targeting of beta(5)RGS7 is a complex process requiring at least RGS domain-mediated interaction with alpha(o) and RGS7 palmitoylation.

Animals↗

PubMed2002

Combining mutations in HIV-1 protease to understand mechanisms of resistance.

HIV-1 develops resistance to protease inhibitors predominantly by selecting mutations in the protease gene. Studies of resistant mutants of HIV-1 protease with single amino acid substitutions have shown a range of independent effects on specificity, inhibition, and stability. Four double mutants, K45I/L90M, K45I/V82S, D30N/V82S, and N88D/L90M were selected for analysis on the basis of observations of increased or decreased stability or enzymatic activity for the respective single mutants. The double mutants were assayed for catalysis, inhibition, and stability. Crystal structures were analyzed for the double mutants at resolutions of 2.2-1.2 A to determine the associated molecular changes. Sequence-dependent changes in protease-inhibitor interactions were observed in the crystal structures. Mutations D30N, K45I, and V82S showed altered interactions with inhibitor residues at P2/P2', P3/P3'/P4/P4', and P1/P1', respectively. One of the conformations of Met90 in K45I/L90M has an unfavorably close contact with the carbonyl oxygen of Asp25, as observed previously in the L90M single mutant. The observed catalytic efficiency and inhibition for the double mutants depended on the specific substrate or inhibitor. In particular, large variation in cleavage of p6(pol)-PR substrate was observed, which is likely to result in defects in the maturation of the protease from the Gag-Pol precursor and hence viral replication. Three of the double mutants showed values for stability that were intermediate between the values observed for the respective single mutants. D30N/V82S mutant showed lower stability than either of the two individual mutations, which is possibly due to concerted changes in the central P2-P2' and S2-S2' sites. The complex effects of combining mutations are discussed.

Amino Acid Substitution↗