RESUMO
Galactinol synthase (GS) was purified 1591-fold with a 3.9% recovery from the cotyledon of kidney bean (Phaseolus vulgaris) by a novel scheme consisting of ammonium sulfate fractionation followed by diethylaminoethyl, Affi-Gel Blue, and UDP-hexanolamine affinity chromatography. The purified enzyme had a specific activity of 8.75 mumol mg-1 min-1, a pH optimum of 7.0, and requirements for manganese ion and DTT. The enzyme exhibited a Km = 0.4 mM for UDP-galactose and a Km = 4.5 mM for myo-inositol. It was identified as a 38-kD peptide that co-purified with a 41- and a 43-kD peptide as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purification to homogeneity was achieved by isolating the 38-kD peptide from the SDS-PAGE gel. To clarify conflicting reports in the literature about the relative molecular mass of purified GS from zucchini leaf (Cucurbita pepo), a similar scheme with modified eluting conditions was used to purify GS from this source. Zucchini leaf GS was purified to homogeneity and identified as a 36-kD peptide on SDS-PAGE. Partial N-terminal sequences of the 38-kD peptide from kidney bean cotyledon and the 36-kD peptide from zucchini leaf were obtained. To facilitate identification of GS during the purification, an assay utilizing thin-layer chromatography and an isotopic analytic imaging scanner was developed.
Assuntos
Fabaceae/enzimologia , Galactosiltransferases/química , Galactosiltransferases/metabolismo , Plantas Medicinais , Verduras/enzimologia , Sequência de Aminoácidos , Cromatografia de Afinidade , Cromatografia DEAE-Celulose , Eletroforese em Gel de Poliacrilamida , Galactosiltransferases/isolamento & purificação , Cinética , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/isolamento & purificação , Folhas de Planta , Sementes , Homologia de Sequência de AminoácidosRESUMO
Health care for rape victims traditionally has been crisis oriented, focused solely upon the victim, and provided either by a gynecologist or psychiatrist. A family practice health team, working in a county hospital, offered rape victims and their families crisis counseling in the emergency room, and follow-up care in a family practice clinic. Follow-up of victims markedly increased from 8% before initiation of the program to 86% afterward. Family members of more than half of the victims seen on follow-up received treatment. A wide range of health needs of both victims and their families, not identified in the emergency room, was identified on follow-up visits. In this way emergency care of the rape victim often became an entry point into the health care system. Rape must be viewed as a family's as well as victim's health problem, often demanding a broader service than can be offered by a gynecologist or psychiatrist alone.