Gelsolin GSN
Background. Gelsolin consists of six homologous domains (G1–G6), each containing Ca-binding sites. Upon occupation of a subset of these sites, gelsolin can cap and depolymerize actin filaments (Nag et al., 2009). Deletion of the extreme COOH terminal region of gelsolin removes a dicarboxylic acid sequence motif similar to a motif found on the NH2 terminus of actin. This deletion results in a loss of Ca2+-requirement for actin monomer binding and thin filament severing activity (Kwiatkowsky, 1989).
Application. The recombinant gelsolin subfragment depolimerizes thin filaments in a calcium-independent manner (Kwiatkowski et al., 1989). Therefore, thin filaments can be removed from intact myofibrils in relaxation buffer, and the acto-myosin independent features, such as the contribution of titin and collagen elasticity can be determined.
Products:
GSN-1: His-tagged gelsolin fragment, 29,364 Dalton
GSN-2: Gelsolin fragment without His-tag, 26,365 Dalton
Sequence of the gelsolin peptide (red tag present in GSN-1 only:
MKHHHHHHPMSDYDIPTTENLYFQGAMAATASRGASQAGAPQGRVPEARP 50
NSMVVEHPEFLKAGKEPGLQIWRVEKFDLVPVPTNLYGDFFTGDAYVILK 100
TVQLRNGNLQYDLHYWLGNECSQDESGAAAIFTVQLDDYLNGRAVQHREV 150
QGFESATFLGYFKSGLKYKKGGVASGFKHVVPNEVVVQRLFQVKGRRVVR 200
ATEVPVSWESFNNGDCFILDLGNNIHQWCGSNSNRYERLKATQVSKGIRD 250
NERSGRAPSARV*
Product analysis by SDS-PAGE and Coomassie stain:
lanes 1,2,3: GSN-1 with His-Tag
lane 4: GSN-2 without His-Tag
References:
Kwiatkowski DJ, Janmey PA, Yin HL. Identification of critical functional and regulatory domains in gelsolin.
J Cell Biol. 1989 May;108(5):1717-26.
Kwiatkowski DJ. Functions of gelsolin: motility, signaling, apoptosis, cancer. Curr Opin Cell Biol. 1999 Feb;11(1):103-8.
Shalini Nag et al. Ca2+ binding by domain 2 plays a critical role in the activation and stabilization of gelsolin Proc. Natl. Acad. Sci. USA 2009 106:13713-13718.