Reconstituted families

Additional strains were placed on marital relations, particularly in reconstituted families where the grandparent s partner was not directly related and did not necessarily share the same acute sense of family obligation to help the forsaken child ... 

It was rare to find families in which the birth parents were still living together (4/24) the most likely scenario was that there had been parental separation and later reconstitution of the family with one, or serial, stepparents. Disagreement between birth parents and step-parents over the child was apt to coalesce around these fault lines and added to the discord ... 

Reconstituted or not, however, the parents interviewed reported high levels of severe and persistent discord between them over how to respond to the child with the drug problem and the costs to the family as a whole. 

The fusion of vesicles with opposite charges appears to be an efficient method to increase the molecular complexity. Suppose all enzymes of the Krebs cycle were distributed into five or six different charged vesicle families (some positively, some negatively charged) - then mixed. Would the Krehs cycle be reconstituted ... 

Although remarkable progress has been made in matching the secondary structure and the function in the various members of the mitochondrial carrier family (del Arco and Satrustegui 2005), satisfying information about the function of potential ADP/ATP transporters can only come from a detailed functional analysis (see later). Several methods are currently available reconstitution in liposomes, in vivo expression, rescue experiments, and functional tests in bacteria (Escherichia coli, Lactococcus lactis). Such tests involve competition experiments, effector and inhibitor studies (Voncken 2001 Voncken et al. 2002a Haferkamp et al. 2002 van der Giezen et al. 2002 Tjaden et al. 2004 Chan et al. 2005 Leroch 2006). 

The idea that the same cofactor species operated in all Mo enzymes originated from a reconstitution assay. In this assay method, the isolated Moco from one enzyme, such as XO, is inserted into a cofactor-free mutant (Nit-1) of nitrate reductase from Neuraspora crassa, where it can reactivate or reconstitute normal nitrate reductase catalytic activity. It is now recognized that the Mo at the active site has many different coordination environments, as has been illustrated for the three Mo families in Fig. 1. In this context, the mutant nitrate reductase assay experiment is interpreted as involving some reprocessing of the inserted molybdenum cofactor from foreign enzymes to obtain the correct form of the cofactor for nitrate reductase catalysis. The Moco designation, if it is to be used, must refer to the family of sites present in Moco enzymes. 

So far, one member of the S2P family has been purified to homogeneity with preservation of proteolytic activity (17). The purification of this protease, the E. coli YaeL/RseP, should allow a more rigorous and direct determination of substrate sequence requirements. Moreover, the ability to purify an I-CLiP and reconstitute activity (in appropriate detergent and usually in the presence of added lipids) is an important step toward studying these unusual membrane-embedded proteases in terms... 

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