Journal: Nature Communications
Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP
Figure Lengend Snippet: MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P
Article Snippet: Fluorescence measurement Nucleotide release kinetics of the fluorescent nucleotide analogue MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and nucleotide binding of MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) were performed in a Perkin Elmer LS55 fluorescence spectrometer instrument (excitation 360 nm, emission 420 nm).
Techniques: Fluorescence, Isolation, Mutagenesis