Constitutive calcium-independent release of Toxoplasma gondii dense granules occurs through the NSF/SNAP/SNARE/Rab machinery.
AuthorsChaturvedi S, Qi H, Coleman D, Rodriguez A, Hanson PI, Striepen B, Roos DS, Joiner KA
JournalJ Biol Chem
PubMed ID9891012
'The signals and the molecular machinery mediating release of dense matrix granules from pathogenic protozoan parasites are unknown. We compared the secretion of the endogenous dense granule marker GRA3 in Toxoplasma gondii with the release of a stably transfected foreign reporter, beta-lactamase, that localizes to parasite dense granules. Both proteins ... More
Optimal configuration of pyrene excimer induced by the excimer-forming two-probe nucleic acid hybridization method.
AuthorsMasuko M, Toyoda S, Suwa M, Mitaku S, Shimazu A, Ohtani H
JournalNucleic Acids Symp Ser
PubMed ID9586010
Excimer-forming two-probe nucleic acid hybridization (ETPH) method with pyrene as a fluorophore enables homogeneous hybridization assays. We examined the effect of linker length between a pyrene residue and a terminal sugar moiety on Tm of hybrids in the presence of 20% dimethylformamide (DMF). The results including those of CD measurements ... More
Nucleic acid hybridization accompanied with excimer formation from two pyrene-labeled probes.
AuthorsEbata K, Masuko M, Ohtani H, Kashiwasake-Jibu M
JournalPhotochem Photobiol
PubMed ID8570721
We developed a novel nucleic acid hybridization method based on excimer formation. We used two different 16-mer oligonucleotide probes that had a combined continuous-sequence run that was complementary to a target 32-mer. Prior to hybridization, the adjacent terminal ends (i.e. the 3'-terminal of one probe and the 5'-terminal of the ... More
Structural mapping of an aggregation nucleation site in a molten globule intermediate.
AuthorsHammarström P, Persson M, Freskgârd PO, Mârtensson LG, Andersson D, Jonsson BH, Carlsson U
JournalJ Biol Chem
PubMed ID10551854
Protein aggregation plays an important role in biotechnology and also causes numerous diseases. Human carbonic anhydrase II is a suitable model protein for studying the mechanism of aggregation. We found that a molten globule state of the enzyme formed aggregates. The intermolecular interactions involved in aggregate formation were localized in ... More
Pyrene excimer fluorescence as a proximity probe for investigation of residual structure in the unfolded state of human carbonic anhydrase II.
AuthorsHammarström P, Kalman B, Jonsson BH, Carlsson U
JournalFEBS Lett
PubMed ID9450551
The excimer fluorescence from two pyrenyl moieties attached to cysteines in human carbonic anhydrase II has been monitored to characterize residual structure retained under strong denaturing conditions. A position in beta-strand 3, N67C, together with the single naturally occurring cysteine 206 in beta-strand 7, were used as attachment sites. The ... More
Structure of Saccharomyces cerevisiae alpha-agglutinin. Evidence for a yeast cell wall protein with multiple immunoglobulin-like domains with atypical disulfides.
alpha-Agglutinin of Saccharomyces cerevisiae is a cell wall-associated protein that mediates cell interaction in mating. Although the mature protein includes about 610 residues, the NH2-terminal half of the protein is sufficient for binding to its ligand a-agglutinin. alpha-Agglutinin20-351, a fully active fragment of the protein, has been purified and analyzed. ... More