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Blood, Vol. 109, Issue 2, 603-609, January 15, 2007
Glycoprotein Ib forms disulfide bonds with 2 glycoprotein Ibß subunits in the resting platelet
Blood Luo et al.
109: 603
Supplemental materials for: Luo et al, Vol 109, Issue 2, 603-609
Files in this Data Supplement:
- Table S1. Product distribution in the thiol-disulfide exchange reactions (PDF, 25.2 KB) -
Each product (identified by peak number) concentration was calculated by integration of the corresponding HPLC peak and was expressed as the percentage of the total peptide concentration. The peak numbers follow the assignment listed in the legend of Figure 3C-D. For the products containing both  and  peptides, 2 percentage numbers are calculated.
- Figure S1. Expression and purification of the Ibα and Ibβ TM peptides (PDF, 1.7 MB) -
(A) SDS gels depicting purification of the Ib (lanes 1-5) and IbSC (lanes 6-10) TM peptides. Lane 1 and 6 indicates whole-cell lysate; lane 2 and 7, isolated inclusion body; lane 3 and 8, His-GST-TM fusion protein eluted from the Ni-agarose column; lane 4 and 9, thrombin cleavage to generate the His-GST fragment and the TM peptide (bottom band); lane 5, purified Ib TM peptide; lane 10, purified IbSC TM peptide. (B) Mass determination of each TM peptide to confirm their identities.
- Figure S2. Surface-expression levels of GP Ibα (black bar) and GP IX (white bar), quantified as relative mean fluorescence, in mutant CHO cells (PDF, 53.4 KB) -
Vectors containing wild-type or mutant cDNAs for GP Ib, Ib, and IX were transiently cotransfected into CHO cells. After 48 to 72 hours, the cells were harvested, stained by anti-Ib antibody AK2 (or SZ2) or anti-IX antibody FMC25, and analyzed by FACS analysis. The mean fluorescence values were normalized as described,21 with IX cells being 100% and cells transfected with empty vectors 0% (not shown). The data are presented as the mean ± SD, calculated from 3 to 4 independent experiments.
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