RESUMO
Recent studies of human populations suggest that the genome consists of chromosome segments that are ancestrally conserved ('haplotype blocks'; refs. 1-3) and have discrete boundaries defined by recombination hot spots. Using publicly available genetic markers, we have constructed a first-generation haplotype map of chromosome 19. As expected for this marker density, approximately one-third of the chromosome is encompassed within haplotype blocks. Evolutionary modeling of the data indicates that recombination hot spots are not required to explain most of the observed blocks, providing that marker ascertainment and the observed marker spacing are considered. In contrast, several long blocks are inconsistent with our evolutionary models, and different mechanisms could explain their origins.
Assuntos
Cromossomos Humanos Par 19/genética , Haplótipos/genética , Recombinação Genética , Alelos , Mapeamento Cromossômico , DNA/genética , Evolução Molecular , Frequência do Gene , Marcadores Genéticos , Humanos , Desequilíbrio de Ligação , Modelos Genéticos , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Heme administration causes inhibition of delta-aminolevulinate synthase (ALAS), best tested in the allylisopropylacetamide (AIA)-treated rat, a model for hepatic porphyrias. Because heme suspended in aqueous media (for injection) is unstable and has adverse effects on coagulation, alternate therapeutic modalities are being explored. The present study tries to answer two questions: 1) are any heme analogs as effective inhibitors of ALAS as heme is; and 2) does heme administration in the form of liposomes increase its effectiveness? None of the liposome compositions tested, even if containing lactosylceramide for preferential hepatocyte uptake, was more effective in inhibiting AIA-induced ALAS activity than heme in buffer. As for the function of the heme analogs, although deuteroheme and heme dimethyl ester proved ineffective, mesoheme and cobalt protoporphyrin were nearly as effective as heme itself, indicating that both hydrophobic side chains in positions 2 and 4 and free propionate groups at 6 and 7 are essential for ALAS inhibition, as is the presence of a central cobalt or iron atom.
Assuntos
5-Aminolevulinato Sintetase/antagonistas & inibidores , Heme/administração & dosagem , Fígado/enzimologia , Metaloporfirinas/farmacologia , Porfirinas/farmacologia , Animais , Transporte Biológico , Soluções Tampão , Lipossomos , Masculino , Ratos , Relação Estrutura-AtividadeRESUMO
As a model for the transport of hemin across biological membranes, sonicated phosphatidylcholine liposomes with incorporated hemin were characterized. The interaction of the hemin liposomes with the heme binding proteins albumin, apomyoglobin, and hemopexin was examined as a function of liposome charge and cholesterol content. In all cases, there was an almost complete transfer of hemin from liposome to protein; a rapid phase and a slow phase were observed for the transfer. For negatively charged liposomes (with 11% dicetyl phosphate), the rapid and slow phases showed observed rates of transfer of ca. 2 and 0.01 s-1, respectively, for all three proteins. The presence of cholesterol in the liposomes decreased the observed rates by a factor of 2, and positively charged liposomes (with 11% stearylamine) showed about one-fifth the observed rates of negatively charged liposomes. The observed rates were independent of protein concentration, indicating that the rate-determining step is hemin efflux from the lipid bilayer. The hemin interaction with the phospholipid bilayer is suggested to be primarily hydrophobic with some electrostatic character. The two phases are suggested to arise from two different populations of hemin within the liposomes and are interpreted as arising from two different orientations of hemin within the bilayer.
