Merck
  • Home
  • Search Results
  • Changes in the Factor VIII C2 domain upon membrane binding determined by hydrogen-deuterium exchange MS.

Changes in the Factor VIII C2 domain upon membrane binding determined by hydrogen-deuterium exchange MS.

The Biochemical journal (2014-05-13)
Dionysios Pantazatos, Christopher R Gessner, Virgil L Woods, Gary E Gilbert
ABSTRACT

Factor VIII enhances the catalytic activity of Factor IXa in a membrane-bound enzyme complex and both proteins are necessary to prevent haemophilia. Tandem lectin-like C domains mediate the membrane binding of Factor VIII and membrane-interactive residues have been identified. However, the available data provide little insight into the dynamic changes that occur upon membrane binding. We used time-based hydrogen-deuterium exchange MS to evaluate the dynamics of FVIII-C2 (Factor VIII C2 domain) alone and when membrane bound. The results confirm the participation of previously identified membrane-interactive loops in the binding mechanism. In addition, they indicate that a long peptide segment, encompassing a membrane-interactive loop and strands of the β-barrel core, is remarkably dynamic prior to membrane binding. The flexibility is reduced following membrane binding. In addition, regions that interact with the A1 and C1 domains have reduced solvent exchange. Thus the isolated C2 domain has extensive flexibility that is subject to stabilization and could be related to interactions between domains as well as between Factor VIII and Factor IXa or Factor X. These results confirm that the proposed membrane-binding loops of the FVIII-C2 interact with the membrane in a manner that leads to protection from solvent exposure.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
盐酸胍 盐酸盐, for molecular biology, ≥99%
Sigma-Aldrich
甲酸, reagent grade, ≥95%
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
胃蛋白酶 来源于猪胃粘膜, powder, ≥250 units/mg solid
Sigma-Aldrich
盐酸胍 盐酸盐, ≥99% (titration), organic base and chaeotropic agent
Sigma-Aldrich
甲酸, ACS reagent, ≥96%
Sigma-Aldrich
胃蛋白酶 来源于猪胃粘膜, lyophilized powder, ≥3,200 units/mg protein
Sigma-Aldrich
甲酸, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥98%
Sigma-Aldrich
盐酸胍 盐酸盐, ≥98%
Sigma-Aldrich
甲酸, puriss., meets analytical specifications of DAC, FCC, 98.0-100%
Sigma-Aldrich
乙酰氯, puriss. p.a., ≥99.0% (T)
Sigma-Aldrich
胃蛋白酶 来源于猪胃粘膜, powder, ≥400 units/mg protein
Sigma-Aldrich
乙酰氯, reagent grade, 98%
SAFC
盐酸胍 盐酸盐
Sigma-Aldrich
异丙 β-D-1-硫代半乳糖苷, ≥99% (TLC)
Sigma-Aldrich
甲酸, ACS reagent, ≥88%
Sigma-Aldrich
胍 盐酸盐 溶液, BioUltra, ~8 M in H2O
Sigma-Aldrich
胍 盐酸盐 溶液
Sigma-Aldrich
乙酰氯, reagent grade, 98%
SAFC
异丙 β-D-1-硫代半乳糖苷
Sigma-Aldrich
异丙基 β-D-硫代半乳糖吡喃糖苷 溶液, ReadyMade IPTG solution for Blue-white screening
胃蛋白酶, powder, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
甲酸, ≥95%, FCC, FG
Sigma-Aldrich
胃蛋白酶 来源于猪胃粘膜, powder, slightly beige, ≥500 U/mg
Sigma-Aldrich
盐酸胍 盐酸盐, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
盐酸胍 盐酸盐, ≥99.0% (AT)
Sigma-Aldrich
甲酸 溶液, BioUltra, 1.0 M in H2O
Sigma-Aldrich
盐酸胍 盐酸盐, anhydrous, free-flowing, Redi-Dri, ≥99%
Sigma-Aldrich
胃蛋白酶 来源于猪胃粘膜, powder, slightly beige, 1200-2400 U/mg
人凝血因子factor浓缩物, European Pharmacopoeia (EP) Reference Standard