Characteristics, importance, and implications of comprehensive drug therapy management.

Ideas and opinions about comprehensive drug therapy management (CDTM) were gathered from an expert panel representing health care practitioners, educators, and consumers. A qualitative method of inquiry, the Delphi technique, was used to aggregate opinions of an expert panel of health care professionals, health care educators, and consumer representatives. The 66 experts who agreed to participate in the study consisted of 10 from pharmacy education, 10 from pharmacy professional associations, 20 from managed care, 19 from primary care, and 7 consumers (public members of health care regulatory and governing boards). Each participant was sent a questionnaire designed to evaluate a preliminary definition of CDTM and gather opinions on related topics. A second questionnaire was sent to the participants to determine the extent of their agreement with each statement generated by the first questionnaire. A third questionnaire was sent, asking participants to reconsider their ratings in light of the group's responses and to choose the most important elements. The response rates were 83% for the first questionnaire, 70% for the second, and 76% for the third. The panel reached a high level of agreement on a definition of CDTM. Panel members also identified many critical aspects of CDTM, including what makes it worthwhile, barriers to and facilitators of engaging in CDTM, key participants, ways in which responsibility for CDTM may be shared, and the competencies needed for CDTM. An expert panel of health care leaders and future leaders, within and outside the pharmacy profession, reached a high level of agreement on a definition of CDTM and identified many critical aspects of this concept.

A novel photoactivatable cross-linker for the functionally-directed region-specific fluorescent labeling of proteins.

A cleavable cross-linking reagent, sulfosuccinimidyl-2(7-azido-4-methylcoumarin-3-acetamido)-ethyl-1,3'- dithiopropionate (SAED), was synthesized for the selective transfer of a coumarin fluorophore from a 'donor' protein to a position near the binding site of an interacting 'target' protein. SAED contains a terminal N-sulfosuccinimidyl ester for conjugation to the donor, a terminal photoactivatable azido-coumarin species for cross-linking with the interacting target, and a central disulfide spacer for the release of the labeled target after cleavage. To evaluate the effectiveness of this labeling reagent, soybean trypsin inhibitor (STI) was derivatized (approximately 0.5 mol/mol) with SAED and then photolyzed in the presence of trypsin. A single fluorescent cross-linked species (6-7 mol% of total STI) was observed by SDS/PAGE and, after reductive cleavage, was shown to be a 1:1 STI-trypsin complex. This complex was not detected without photolysis or with an inactivated cross-linker. Importantly, complex formation was inhibited by an excess of unmodified STI and prevented by substitution of a non-interacting protein for trypsin. Cleavage of the cross-linked complex revealed that the trypsin, but not the STI, was fluorescent; the uncomplexed trypsin fraction remained unlabeled. These results demonstrated the specificity of the labeling of trypsin by fluorescent-transfer cross-linking with SAED. An efficiency of about 15% for this cross-linking mediated labeling of trypsin was calculated. The short cross-linking span of SAED (less than or equal to 1.8 nm) strictly limited the labeling to the vicinity of the contact region of trypsin with STI. Thus, this novel cross-linker permits the region-specific targeting of a fluorophore near a functionally important binding site.

Amino acids bracketing the predicted transmembrane domains of membrane proteins.

The cell membrane is a complex mixture of several classes of biomolecules but amino acids and lipids are the main constituents. For this reason we are establishing a data base of transmembrane proteins with the intent of using the data base to identify interfacial peptide sequences useful for studying protein-lipid interactions at membrane interfaces. Our present intention is to characterize transmembrane peptides and amino acids found near the membrane interface. A data base containing only signal peptides is available (G. von Heijne, Prot. Seq. Data Anal. 1:41-42, 1987).