INVESTIGADORES
VIZOSO PINTO Maria Guadalupe
capítulos de libros
Título:
LuMPIS: luciferase-based MBP-pull-down protein interaction screening system
Autor/es:
VIZOSO PINTO, M. G.; BAIKER, A.
Libro:
Methods in Molecular Biology 815 - Functional Genomics: Methods and Protocols
Editorial:
Humana Press
Referencias:
Año: 2012; p. 265 - 275
Resumen:
Analyzing the putative interaction partners of an individual protein is
one approach to elucidate its function. In the LuMPIS protocol, bait and
prey proteins are expressed with N-terminal maltose binding protein
(MBP)- and eGFP-luciferase (eGFP-luc) tags, respectively. Positive
protein-protein interactions (PPIs) can be detected after pull-down of
the MBP-tagged prey protein using amylose beads followed by the
bioluminescence detection of the bound eGFP-luc-tagged bait protein. The
LuMPIS technology offers the following advantages: the PPIs are
detected in the mammalian cell context, the use of two long protein tags
(i.e., MBP and eGFP-luc) increases the expression levels of genes whose
gene expression levels are known to be frequently impaired, the use of
amylose beads for pull-down is much more economic as compared to
sepharose beads in combination with monoclonal antibodies and finally,
the use of an eGFP-luc-tag enables the qualitative control of
transfection efficiencies by fluorescence microscopy prior to starting
the assay.
