Molecular modelling of human complement component C3 and its fragments by solution scattering

Perkins, Stephen J.; Sim, Robert B.

doi:10.1111/j.1432-1033.1986.tb09652.x

Cited by 211 publications

(298 citation statements)

References 66 publications

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“…Concentrations c were calculated from calculated absorbance measurements at 280 nm using absorption coefficients (1%, 1 cm) of 12.8, 15.6 and 13.7 for FVIIa, sTF and their complex, respectively [15]. Sequences and carbohydrate contents were obtained from refs.…”

Section: Preparations Of Factor Viia Tissue Factor and Their Complexmentioning

confidence: 99%

Factor VIIa and the extracellular domains of human tissue factor form a compact complex: A study by X‐ray and neutron solution scattering

et al. 1995

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The four-domain structure of human factor Vlla and the two-domain structure of tissue factor form a tight complex to initiate blood coagulation. By solution scattering, the mean X-ray and neutron radii of gyration Re (which determine macromolecular elongation) were found to be 3.25 nm, 2.13 nm and 3.14 nm (+ 0.13 rim) for factor VIIa, the extracellular region of tissue factor and their complex in that order. The mean cross-sectional radii of gyration Rxs were 1.33 rim, 0.56 nm and 1.42 nm ( + 0.13 nm) in that order. The mean lengths were 10.3 nm, 7.7 nm and 10.2 nm in that order. The data show that, in solution, the free proteins have extended domain structures, and the complex is formed by a compact side-by-side alignment of the two proteins along their long axes. The high binding affinity of tissue factor for factor Vlla may thus be accounted for by the occurrence of many intermolecular contacts in the complex.

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Section: Preparations Of Factor Viia Tissue Factor and Their Complexmentioning

confidence: 99%

Factor VIIa and the extracellular domains of human tissue factor form a compact complex: A study by X‐ray and neutron solution scattering

et al. 1995

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“…The density and viscosity were calculated using polynomial equations and tables of coefficients (44). The partial specific volume of TTR (0.734 cm 3 /g) was estimated on the basis of the partial specific volumes of the component amino acid residues (45)(46)(47).…”

Section: Methodsmentioning

confidence: 99%

Characterization of the Transthyretin Acid Denaturation Pathways by Analytical Ultracentrifugation: Implications for Wild-Type, V30M, and L55P Amyloid Fibril Formation

1998

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Analytical ultracentrifugation methods were utilized to further characterize the acid denaturation pathways of wild-type, V30M, and L55P transthyretin (TTR) that generate intermediates leading to amyloid fibril formation and possibly the diseases senile systemic amyloidosis and familial amyloid polyneuropathy. Equilibrium and velocity methods were employed herein to characterize the TTR quaternary structural requirements for amyloid fibril formation. From neutral to slightly acidic conditions (pH 7.5-5.1), wildtype transthyretin (0.2-0.3 mg/mL, 100 mM KCl, 37°C) exists as a tetramer and is incapable of fibril formation. Under more acidic conditions (pH 5 to 3.9), tetrameric wild-type TTR slowly dissociates to a monomer having an alternatively folded tertiary structure(s) that self-assembles at physiological concentration (0.2 mg/mL) into a ladder of quaternary structural intermediates of increasing molecular weight. These intermediates appear to be on the pathway of amyloid fibril formation, since they ultimately disappear when amyloid fibrils are observed. The V30M and L55P TTR variants exhibit similar acid denaturation pathways, with the exception that dissociation of the tetramer to the monomeric amyloidogenic intermediate occurs at a higher pH and to a much greater extent, allowing the quaternary structural intermediates to be readily observed by velocity methods. Partial denaturation and assembly of the monomeric amyloidogenic intermediate(s) occur at pH 5.4 for V30M and L55P TTR over a 72 h period, during which wild-type TTR maintains its normal tetrameric three-dimensional structure. Interestingly, the L55P and V30M familial amyloid polyneuropathy (FAP) associated variants form amyloid protofilaments at pH 7.5 (37°C) after several weeks of incubation, suggesting that the activation barriers for TTR tetramer dissociation to the monomeric amyloidogenic intermediate are much lower for the FAP variants relative to wild-type TTR, which does not form amyloid or amyloid protofilaments under these conditions. This study establishes the key role of the monomeric amyloidogenic intermediate and its selfassembly into a ladder of quaternary structural intermediates for the formation of wild-type, V30M, and L55P transthyretin amyloid fibrils.

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“…The density and viscosity were calculated using polynomial equations and tables of coefficients (36). The partial specific volume of WT-and ∆TM-FAAH (0.738 cm 3 / g) was estimated on the basis of the partial specific volumes of the component amino acid residues (37)(38)(39).…”

Section: Methodsmentioning

confidence: 99%

Comparative Characterization of a Wild Type and Transmembrane Domain-Deleted Fatty Acid Amide Hydrolase: Identification of the Transmembrane Domain as a Site for Oligomerization

et al. 1998

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Fatty acid amide hydrolase (FAAH) is an integral membrane protein responsible for the hydrolysis of a number of primary and secondary fatty acid amides, including the neuromodulatory compounds anandamide and oleamide. Analysis of FAAH's primary sequence reveals the presence of a single predicted transmembrane domain at the extreme N-terminus of the enzyme. A mutant form of the rat FAAH protein lacking this N-terminal transmembrane domain (∆TM-FAAH) was generated and, like wild type FAAH (WT-FAAH), was found to be tightly associated with membranes when expressed in COS-7 cells. Recombinant forms of WT-and ∆TM-FAAH expressed and purified from Escherichia coli exhibited essentially identical enzymatic properties which were also similar to those of the native enzyme from rat liver. Analysis of the oligomerization states of WT-and ∆TM-FAAH by chemical cross-linking, sedimentation velocity analytical ultracentrifugation, and size exclusion chromatography indicated that both enzymes were oligomeric when membrane-bound and after solubilization. However, WT-FAAH consistently behaved as a larger oligomer than ∆TM-FAAH. Additionally, SDS-PAGE analysis of the recombinant proteins identified the presence of SDS-resistant oligomers for WT-FAAH, but not for ∆TM-FAAH. Self-association through FAAH's transmembrane domain was further demonstrated by a FAAH transmembrane domain-GST fusion protein which formed SDS-resistant dimers and large oligomeric assemblies in solution.

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Molecular modelling of human complement component C3 and its fragments by solution scattering

Cited by 211 publications

References 66 publications

Factor VIIa and the extracellular domains of human tissue factor form a compact complex: A study by X‐ray and neutron solution scattering

Factor VIIa and the extracellular domains of human tissue factor form a compact complex: A study by X‐ray and neutron solution scattering

Characterization of the Transthyretin Acid Denaturation Pathways by Analytical Ultracentrifugation: Implications for Wild-Type, V30M, and L55P Amyloid Fibril Formation

Comparative Characterization of a Wild Type and Transmembrane Domain-Deleted Fatty Acid Amide Hydrolase: Identification of the Transmembrane Domain as a Site for Oligomerization

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