Mapping the Binding Region on the Low Density Lipoprotein Receptor for Blood Coagulation Factor VIII

Kurasawa, James H; Shestopal, Svetlana A; Karnaukhova, Elena; Struble, Evi; Lee, Timothy K.; Sarafanov, Andrey G.

doi:10.1074/jbc.m113.468108

Cited by 15 publications

(50 citation statements)

References 67 publications

(79 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Ldlr -/- mice may also develop endothelial disturbances due to the LDL receptor deficiency itself. This assumption can be corroborated by data showing an increased inactivation of H 2 O 2 (an endothelial relaxation factor in Ldlr -/- mice) [ 25 , 26 ] and by the fact that the LDL receptor serves as a binding site for the blood coagulation factor VIII, thus mediating its clearance from the circulation [ 27 ]. Taken together, it is very likely that hypercholesterolemia-dependent and -independent mechanisms contribute to CSVD development in Ldlr -/- mice.…”

Section: Discussionmentioning

confidence: 89%

Hypercholesterolemia induced cerebral small vessel disease

et al. 2017

View full text Add to dashboard Cite

BackgroundWhile hypercholesterolemia plays a causative role for the development of ischemic stroke in large vessels, its significance for cerebral small vessel disease (CSVD) remains unclear. We thus aimed to understand the detailed relationship between hypercholesterolemia and CSVD using the well described Ldlr-/- mouse model.MethodsWe used Ldlr-/- mice (n = 16) and wild-type (WT) mice (n = 15) at the age of 6 and 12 months. Ldlr-/- mice develop high plasma cholesterol levels following a high fat diet. We analyzed cerebral capillaries and arterioles for intravascular erythrocyte accumulations, thrombotic vessel occlusions, blood-brain barrier (BBB) dysfunction and microbleeds.ResultsWe found a significant increase in the number of erythrocyte stases in 6 months old Ldlr-/- mice compared to all other groups (P < 0.05). Ldlr-/- animals aged 12 months showed the highest number of thrombotic occlusions while in WT animals hardly any occlusions could be observed (P < 0.001). Compared to WT mice, Ldlr-/- mice did not display significant gray matter BBB breakdown. Microhemorrhages were observed in one Ldlr-/- mouse that was 6 months old. Results did not differ when considering subcortical and cortical regions.ConclusionsIn Ldlr-/- mice, hypercholesterolemia is related to a thrombotic CSVD phenotype, which is different from hypertension-related CSVD that associates with a hemorrhagic CSVD phenotype. Our data demonstrate a relationship between hypercholesterolemia and the development of CSVD. Ldlr-/- mice appear to be an adequate animal model for research into CSVD.

show abstract

Section: Discussionmentioning

confidence: 89%

Hypercholesterolemia induced cerebral small vessel disease

et al. 2017

View full text Add to dashboard Cite

show abstract

“…We are conscious that our study has some limitations, and among these the different FVIII concentrates used in the patients. Concerning the FL and BDD FVIII products, the LDLR binds an FVIII epitope involving multiple lysine residues located in the A3, C1, and C2 domains, and thus outside the B domain. Finding that the genetic associations were substantially maintained by analysis of FVIII concentrates, would suggest that the genetic components on PKs, both ABO ‐ and LDLR ‐related, exert quite robust effects.…”

Section: Discussionmentioning

confidence: 99%

“…Although HA patients genetically experience low FVIII levels, the treatment with high concentrations of FVIII transiently exposes HA patients to high peaks of circulating FVIII, comparable to those of patients with cardiovascular disease. This could favor the interaction between FVIII and the LDLR, mediated by four adjacent complement type repeats (CR 2‐5) forming a binding site for FVIII with noticeable affinity (40‐90 nmol/L) however, this affinity could be potentially insufficient to bind efficiently the low concentration (0.3 nmol/L) of FVIII in plasma. Thus, significant binding between FVIII and LDLR would require, in addition to increased FVIII levels present in cardiovascular disease or after infusion in HA patients, heparan‐sulfate like proteoglycans, whose role for FVIII binding is still undefined.…”

Section: Discussionmentioning

confidence: 99%

“…Among these, the LDLR, the key member of the family of endocytic receptors, represents a noticeable candidate because of biochemical, animal model‐related, and epidemiological evidences. The region of complement‐type repeats 2‐5 in LDLR was defined as the binding site for FVIII and LDLR has been involved in FVIII clearance in animal models, in which LDLR deficiency prolonged the half‐life of FVIII. In humans, we have reported that single‐nucleotide polymorphisms in the LDLR gene independently associated with high plasma levels of FVIII in patients with coronary artery disease (CAD) .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Functional polymorphisms in the LDLR and pharmacokinetics of Factor VIII concentrates

Lunghi

Bernardi

Martinelli

et al. 2019

Journal of Thrombosis and Haemostasis

View full text Add to dashboard Cite

Background: Optimization of factor VIII (FVIII) infusion in hemophilia A would benefit from identification of FVIII pharmacokinetics (PK) determinants. The low-density lipoprotein receptor (LDLR) contains an FVIII-binding site and might influence FVIII clearance. Consistently, LDLR polymorphisms have been associated with FVIII levels.Objective: To investigate the relationships between individual FVIII PK and functional LDLR polymorphisms. Patients/Methods: Thirty-three hemophilia A patients (FVIII coagulant activity [FVIII:C] ≤2 IU/dL) without inhibitors underwent 85 FVIII single-dose (21.4-51.8 IU/ kg) PKs with different FVIII concentrates. Twenty patients underwent repeated PKs (2-6). FVIII:C measured up to 72 hours was analyzed by two-compartment model. Parameters were evaluated in relation to F8 mutations, ABO blood-group and LDLR genotypes.Results: F8 mutation types were not associated with PK parameters. ABO and LDLR c.1773C/T polymorphism were associated with Alpha, Alpha HL, CLD2, K1-2, and K2-1 parameters, suggesting an influence on the FVIII initial distribution phase.Regression analysis showed an independent association of both ABO and LDLR c.1773C/T with PK parameters (Alpha, β-coefficient −0.311 vs 0.348; CLD2, β-coefficient −0.335 vs 0.318), giving rise to an additive effect in subjects stratified by combined phenotypes. Differently, the LDLR c.81C/T was associated with FVIII clearance and volume of distribution at steady state, which could be related to distinct effects of polymorphisms, potentially linked to LDLR intracellular distribution and FVIII binding behavior. Conclusions:With the limitation of different FVIII concentrates and low number of patients, our data show plausible associations of LDLR polymorphisms with FVIII PK parameters, thus supporting their investigation as candidate functional determinants of FVIII PK. K E Y W O R D SABO blood-group, factor VIII, hemophilia A, LDL-receptor polymorphism, pharmacokinetics | 1289 LUNGHI et aL.

show abstract

“…Indeed, (i) disruption of the RAP gene in mouse model resulted in impairing the expression of these receptors, except LDLR ( 21 , 22 ); (ii) cotransfection of the RAP gene in cell culture facilitated expression of recombinant vLDLR and LRP1, but not LDLR ( 20 ), and (iii) coexpression of RAP and the LRP1 exodomain in cell culture resulted in increase of the latter's yield ( 19 ). At the same time, expression of recombinant CR fragments of LRP1 and LDLR in cell culture yielded relatively low amounts of correctly folded proteins ( 28 , 32 , 33 ) indicating requirement of a folding factor. Notably, the affinity of RAP for LDLR was found to be similar ( 32 ) or weaker ( 34 ) than that for LRP1.…”

mentioning

confidence: 99%

Molecular chaperone RAP interacts with LRP1 in a dynamic bivalent mode and enhances folding of ligand-binding regions of other LDLR family receptors

Marakasova

Olivares

Karnaukhova

et al. 2021

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The low-density lipoprotein receptor (LDLR) family of receptors are cell-surface receptors that internalize numerous ligands and play crucial role in various processes, such as lipoprotein metabolism, hemostasis, fetal development, etc. Previously, receptor-associated protein (RAP) was described as a molecular chaperone for LDLR-related protein 1 (LRP1), a prominent member of the LDLR family. We aimed to verify this role of RAP for LRP1 and two other LDLR family receptors, LDLR and vLDLR, and to investigate the mechanisms of respective interactions using a cell culture model system, purified system, and in silico modelling. Upon coexpression of RAP with clusters of the ligand-binding complement repeats (CRs) of the receptors in secreted form in insect cells culture, the isolated proteins had increased yield, enhanced folding, and improved binding properties compared with proteins expressed without RAP, as determined by circular dichroism and surface plasmon resonance. Within LRP1 CR-clusters II and IV, we identified multiple sites comprised of adjacent CR doublets, which provide alternative bivalent binding combinations with specific pairs of lysines on RAP. Mutational analysis of these lysines within each of isolated RAP D1/D2 and D3 domains having high affinity to LRP1 and of conserved tryptophans on selected CR-doublets of LRP1, as well as in silico docking of a model LRP1 CR-triplet with RAP, indicated a universal role for these residues in interaction of RAP and LRP1. Consequently, we propose a new model of RAP interaction with LDLR family receptors based on switching of the bivalent contacts between molecules over time in a dynamic mode.

show abstract

Mapping the Binding Region on the Low Density Lipoprotein Receptor for Blood Coagulation Factor VIII

Cited by 15 publications

References 67 publications

Hypercholesterolemia induced cerebral small vessel disease

Hypercholesterolemia induced cerebral small vessel disease

Functional polymorphisms in the LDLR and pharmacokinetics of Factor VIII concentrates

Molecular chaperone RAP interacts with LRP1 in a dynamic bivalent mode and enhances folding of ligand-binding regions of other LDLR family receptors

Contact Info

Product

Resources

About