Design of a Molecular Beacon DNA Probe with Two Fluorophores This work was partially supported by the Office of Naval Research Young Investigator Award N00014-98-1-0621 and by an NSF Career Award (CHE-9733650). We would like to thank Dr. Richard Hogrefe, Paul Imperial, Kelly Christianson, and the Trilink Oligonucleotide Synthesis group for producing the molecular beacons used in this study.
AuthorsZhang P, Beck T, Tan W
JournalAngew Chem Int Ed Engl
PubMed ID11180338
The pro domain of beta-secretase does not confer strict zymogen-like properties but does assist proper folding of the protease domain.
AuthorsShi XP, Chen E, Yin KC, Na S, Garsky VM, Lai MT, Li YM, Platchek M, Register RB, Sardana MK, Tang MJ, Thiebeau J, Wood T, Shafer JA, Gardell SJ
JournalJ Biol Chem
PubMed ID11266439
'beta-Secretase (BACE) is a membrane-bound aspartyl protease that cleaves the amyloid precursor protein to generate the N terminus of the amyloid beta peptide. BACE is expressed as a precursor protein containing Pre, Pro, protease, transmembrane, and cytosolic domains. A soluble BACE derivative (PreProBACE460) that is truncated between the protease and ... More
Fluorescent labeling of cell-free synthesized proteins by incorporation of fluorophore-conjugated nonnatural amino acids.
AuthorsKang SH, Jun SY, Kim DM
JournalAnal Biochem
PubMed ID17113028
'Although fluorescent dyes, such as fluorescein derivatives, have bulky and complex structures, nonnatural amino acids carrying these fluorescein derivatives are acceptable by the Escherichia coli ribosome and are useful for the cotranslational fluorescent labeling of cell-free synthesized proteins. Surprisingly, the incorporation efficiency of nonnatural amino acids carrying fluorescein derivatives into ... More
Custom fluorescent-nucleotide synthesis as an alternative method for nucleic acid labeling.
AuthorsHenegariu O, Bray-Ward P, Ward DC
JournalNat Biotechnol
PubMed ID10700155
The variety of potentially useful dyes or haptenes available for fluorescent nucleic acid hybridization assays is far greater than what can be obtained from commercial sources. Since this diversity could be useful in many laboratory applications, we have developed a simple and inexpensive procedure for preparing nonpurified labeled nucleotides, for ... More
Identification and mapping of protein-protein interactions between gp32 and gp59 by cross-linking.
AuthorsIshmael FT, Alley SC, Benkovic SJ
JournalJ Biol Chem
PubMed ID11309384
The bacteriophage T4 59 protein (gp59) plays a vital role in recombination and replication by promoting the assembly of the gene 41 helicase (gp41) onto DNA, thus enabling replication as well as strand exchange in recombination. Loading of the helicase onto gp32 (the T4 single strand binding protein)-coated single-stranded DNA ... More
Absence of IFN-gamma or IL-12 has different effects on experimental myasthenia gravis in C57BL/6 mice.
Immunization with acetylcholine receptor (AChR) causes experimental myasthenia gravis (EMG). Th1 cells facilitate EMG development. IFN-gamma and IL-12 induce Th1 responses: we investigated whether these cytokines are necessary for EMG development. We immunized wild-type (WT) C57BL/6 mice and IFN-gamma and IL-12 knockout mutants (IFN-gamma-/-, IL-12-/-) with Torpedo AChR (TAChR). WT ... More
Double-modification of lectin using two distinct chemistries for fluorescent ratiometric sensing and imaging saccharides in test tube or in cell.
AuthorsNakata E, Koshi Y, Koga E, Katayama Y, Hamachi I
JournalJ Am Chem Soc
PubMed ID16173755
The site-selective incorporation of two different fluorophores into a naturally occurring protein (lectin, a sugar-binding protein) has been successfully carried out using two distinct orthogonal chemical methods. By post-photoaffinity labeling modification, Con A, a glucose- and mannose-selective lectin, was modified with fluorescein in the proximity of the sugar binding site ... More