Critical residues and motifs for homodimerization of the first transmembrane domain of the plasma membrane glycoprotein CD36.

The plasma transmembrane (TM) glycoprotein CD36 is critically involved in many essential signaling processes, especially the binding/uptake of long-chain fatty acids and oxidized low-density lipoproteins. The association of CD36 potentially activates cytosolic protein tyrosine kinases that are thought to associate with the C-terminal cytoplasmic tail of CD36. To understand the mechanisms by which CD36 mediates ligand binding and signal transduction, we have characterized the homo-oligomeric interaction of CD36 TM domains in membrane environments and with molecular dynamics (MD) simulations. Analysis of pyrene- and coumarin-labeled TM1 peptides in SDS by FRET confirmed the homodimerization of the CD36 TM1 peptide. Homodimerization assays of CD36 TM domains with the TOXCAT technique showed that its first TM (TM1) domain, but not the second TM (TM2) domain, could homodimerize in a cell membrane. Small-residue, site-specific mutation scanning revealed that the CD36 TM1 dimerization is mediated by the conserved small residues Gly12, Gly16, Ala20, and Gly23 Furthermore, molecular dynamics (MD) simulation studies demonstrated that CD36 TM1 exhibited a switching dimerization with two right-handed packing modes driven by the 12GXXXGXXXA20 and 20AXXG23 motifs, and the mutational effect of G16I and G23I revealed these representative conformations of CD36 TM1. This packing switch pattern of CD36 TM1 homodimer was further examined and confirmed by FRET analysis of monobromobimane (mBBr)-labeled CD36 TM1 peptides. Overall, this work provides a structural basis for understanding the role of TM association in regulating signal transduction via CD36.

Complementarity in Requirements Tracing.

Complementarity between activities reveals that doing any one of them increases the returns to doing the others. In other words, complementarity leads to the synergistic effect that the whole is greater than the sum of its parts. Identifying and exploiting complementarity can benefit many cybernetic activities, where human-machine interactions are inherent and dominant. One such activity is requirements tracing that helps stakeholders to track the status of their goals. Although various kinds of support for human analysts in requirements tracing have been proposed, little is known about the nature of complementarity when different tracing practices are involved. In this paper, we explore the role of complementarity by considering together the tagging-to-trace (T2T) and learning-to-trace (L2T) activities. We present a novel approach to examining which T2T and L2T practices enhance the qualities of each other. Our approach also uncovers the environmental factors which the complementarity is sensitive to. Applying our approach to the logs of 140 analyst-tracing units offers operational insights into the rigorous detection of complementarity and shows the importance of understanding the cybernetic conditions under which the requirements tracing practices may in fact be complementary.