Transcript Powerpoint - Blood Journal

Protein S Gla-domain mutations causing impaired Ca2+induced phospholipid binding and severe functional
protein S deficiency
by Suely M. Rezende, David A. Lane, Blandine Mille-Baker, Michel M. Samama,
Jacqueline Conard, and Rachel E. Simmonds
Blood
Volume 100(8):2812-2819
October 15, 2002
©2002 by American Society of Hematology
Pedigree of the family under investigation.Males are shown as squares, females as circles, and
deceased family members are crossed through.
Suely M. Rezende et al. Blood 2002;100:2812-2819
©2002 by American Society of Hematology
Relative transient expression levels of recombinant wild-type and variant protein S in COS-1
cells.Mean, relative concentration of protein S expressed after correction for transfection
efficiency.
Suely M. Rezende et al. Blood 2002;100:2812-2819
©2002 by American Society of Hematology
Western blotting of wild-type protein S and variants expressed in HEK293 cells.SDS-PAGE of 30
ng protein S under reducing conditions and immunoblotting with a polyclonal protein S antibody
were performed as described in the text.
Suely M. Rezende et al. Blood 2002;100:2812-2819
©2002 by American Society of Hematology
Enhancement of APC-dependent factor Va inactivation by wild-type protein S and variants.A
range of recombinant protein S concentrations (0 to 20 nM) was incubated with 0.1 nM APC, 3
nM factor Va, and 25 μM phospholipid vesicles for 2 minutes at 37°C.
Suely M. Rezende et al. Blood 2002;100:2812-2819
©2002 by American Society of Hematology
Binding of recombinant human protein S to phospholipid vesicles.PE/PC/PS 40:40:20
phospholipid vesicles were coated onto the wells of microtiter plates and blocked with 3%
(wt/vol) BSA. (A) Dialyzed recombinant protein S (0 to 26 nM) was incubated in the we...
Suely M. Rezende et al. Blood 2002;100:2812-2819
©2002 by American Society of Hematology
Interaction of recombinant human protein S with Ca2+-dependent monoclonal
antibodies.Antibodies were coated into the wells of microtiter plates at 10 μg/mL and blocked
with casein.
Suely M. Rezende et al. Blood 2002;100:2812-2819
©2002 by American Society of Hematology
Predicted effect of the Gly11Asp mutation on the conformation of the protein S Gla domain.The
Gla domain of protein S shown was derived by homology modeling and refinement.36 In order to
predict the effects of the Gly11Asp amino acid substitution on protein...
Suely M. Rezende et al. Blood 2002;100:2812-2819
©2002 by American Society of Hematology