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Dynein is a minus end-directed microtubule motor that serves multiple cellular functions. We have performed a fine mapping of the 8 kDa dynein light chain (LC8) binding sites throughout the development of a library of consecutive synthetic dodecapeptides covering the amino acid sequences of the various proteins known to interact with this dynein member according to the yeast two hybrid system. Two different consensus sequences were identified: GIQVD present in nNOS, in DNA cytosine methyl transferase and also in GKAP, where it is present twice in the protein sequence. The other LC8 binding motif is KSTQT, present in Bim, dynein heavy chain, Kid-1, protein 4 and also in swallow. Interestingly, this KSTQT motif is also present in several viruses known to associate with microtubules during retrograde transport from the plasma membrane to the nucleus during viral infection.

Apoptotic protease activating factor-1 (Apaf-1) is an adaptor molecule essential for caspase-9 activation. Subcellular analysis of Apaf-1 in NIH-3T3 fibroblasts and the immature murine B cell lymphoma WEHI-231 indicates that Apaf-1 is localized in the Golgi apparatus and cytoplasm. Bcl-2 overexpression in WEHI-231 cells disrupts Apaf-1 localization in Golgi, causing a perinuclear Apaf-1 redistribution. Bcl-2 overexpression in NIH-3T3 fibroblasts however does not cause similar Apaf-1 redistribution, suggesting that cell type factors are involved in the redistribution process. The ability of Bcl-2 to modify Apaf-1 subcellular localization is not explained by direct interaction between Apaf-1 and Bcl-2. These data may help to clarify the anti-apoptotic Bcl-2 function.

A 20-amino acid synthetic peptide from the N-terminal region of gamma3 avenin yields a surprisingly strong reactivity with anti-gliadin antibodies (AGA) of coeliac sera, comparable to that of a gliadin extract. In contrast, a low reactivity is observed with five similar peptides derived from alpha-gliadin, gamma70 and omega1 secalins. Circular dichroism studies of these peptides show that the avenin peptide displays the highest beta-turn content (30%), while other peptides yield much lower values. In agreement with circular dichroism data, nuclear magnetic resonance data point to the presence of a beta-turn in the avenin peptide DPSEQ segment, a sequence with a high statistical beta-turn preference. A strong linear dependence between AGA reactivity and beta-turn content was observed for these peptides, indicating for the first time a role of beta-turn motifs in anti-gliadin antibodies recognition in coeliac disease. This suggests that circulating AGA in coeliac patients comprise not only linear but also conformational antibodies against beta-turn motifs. Polyclonal antibodies raised against the avenin peptide containing beta-turn motifs react by immunoblotting with all gliadin, hordein and secalin proteins, which are rich in beta-turn conformations, despite that their primary structures are unrelated to that of the peptide.