Kinetics and Thermodynamics of Annexin A1 Binding to Solid-Supported Membranes:  A QCM Study

Kastl, Katja; Ross, Michaela; Gerke, Volker; Steinem, Claudia

doi:10.1021/bi025951z

Cited by 67 publications

(72 citation statements)

References 55 publications

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“…As expected, A1-mβG exhibited higher K d values than A5-mβG. However, A1-mβG displayed lower K d values for cultured cells expressing phosphatidylserine than previously reported for A1 alone binding to immobilized lipid bilayers containing phosphatidylserine,~39 nM (Kastl et al, 2002); there are no previous reports of A1 dissociation constants for cultured cells. We have previously shown that annexin A5 fusion proteins will not bind to normal endothelial cell controls Krais et al, 2013), and we here extend this finding to A1 fusion proteins by demonstrating that A1-mβG and A5-mβG bind only to cells expressing phosphatidylserine in a calcium-dependent manner with no detectable specific binding to normal controls lacking the phosphatidylserine target.…”

Section: Discussionsupporting

confidence: 62%

Annexin-directed β-glucuronidase for the targeted treatment of solid tumors

Guillen

Ruben

Virani

et al. 2016

Protein Engineering, Design and Selection

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Enzyme prodrug therapy has the potential to remedy the lack of selectivity associated with the systemic administration of chemotherapy. However, most current systems are immunogenic and constrained to a monotherapeutic approach. We developed a new class of fusion proteins centered about the human enzyme β-glucuronidase (βG), capable of converting several innocuous prodrugs into chemotherapeutics. We targeted βG to phosphatidylserine on tumor cells, tumor vasculature and metastases via annexin A1/A5. Phosphatidylserine shows promise as a universal marker for solid tumors and allows for tumor type-independent targeting. To create fusion proteins, human annexin A1/A5 was genetically fused to the activity-enhancing 16a3 mutant of human βG, expressed in chemically defined, fed-batch suspension culture, and chromatographically purified. All fusion constructs achieved >95% purity with yields up to 740 μg/l. Fusion proteins displayed cancer selective cell-surface binding with cell line-dependent binding stability. One fusion protein in combination with the prodrug SN-38 glucuronide was as effective as the drug SN-38 on Panc-1 pancreatic cancer cells and HAAE-1 endothelial cells, and demonstrated efficacy against MCF-7 breast cancer cells. βG fusion proteins effectively enable localized combination therapy that can be tailored to each patient via prodrug selection, with promising clinical potential based on their near fully human design.

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Section: Discussionsupporting

confidence: 62%

Annexin-directed β-glucuronidase for the targeted treatment of solid tumors

Guillen

Ruben

Virani

et al. 2016

Protein Engineering, Design and Selection

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“…Typical annexins carry 4 similar domains defined by 5 α-helices, and each domain contains a characteristic type 2 calcium-binding motif [GXGTD-[ X ] 37 -(D/E)]. Most of the evolutionarily conserved annexins can interact with PS [Kastl et al, 2002;Rosenbaum et al, 2011], and during skin injury they can bind at the site of tissue damage to suppress the binding and activation of coagulation factors at the cell surface but also promote repair [Bouter et al, 2011].…”

Section: Introductionmentioning

confidence: 99%

Skin Wound Repair Is Not Altered in the Absence of Endogenous AnxA1 or AnxA5, but Pharmacological Concentrations of AnxA4 and AnxA5 Inhibit Wound Hemostasis

Kreft¹,

Klatt

Strassburger

et al. 2016

Cells Tissues Organs

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Skin injury induces the cell surface exposure of phosphatidylserine (PS) on damaged and dying cells to activate coagulation and repair processes. Annexins can bind to PS and may modulate the healing response. Here, we determine the relevance of annexins for skin wound healing using AnxA1- and AnxA5-deficient mice and recombinant annexins with distinct PS binding properties. Wound inflammation, closure and the formation of granulation tissue were not altered in AnxA1- or AnxA5-deficient mice or after increasing AnxA5 serum concentrations (100 nM) in wild-type mice. Increased serum concentrations (1 µM) of AnxA5 induced massive bleeding, but wound hemostasis was not delayed by AnxA1. Both annexins interact with PS, but only AnxA5 can form 2-dimensional (2D) arrays on the cell surface. The injection of an AnxA5 mutant that binds to PS but lacks the ability of 2D array formation failed to induce bleeding. 2D lattice-forming AnxA4, with high affinity to PS also caused bleeding, while hemostasis was not affected by AnxA8 with low affinity or the AnxA8 mutant with medium affinity for PS and the lack of 2D formation. Increased concentrations of AnxA4 and AnxA5 also delayed coagulation pathway activation in vitro. This effect was attenuated for the AnxA5 mutant as well as for AnxA1 and AnxA8. In conclusion, endogenous AnxA1 and AnxA5 are dispensable for wound hemostasis and repair, but pharmacologically excessive concentrations of AnxA4 and AnxA5 inhibit hemostasis in skin wounds.

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“…It was adsorbed on a POPC:(palmitoyloleoylphosphatidylserine = POPS) (4:1) lipid monolayer assembled by vesicle fusion on an octanethiol monolayer tethered to the gold surface of a QCM. 91 Adsorption of annexin A1 to this hybrid bilayer in the presence of Ca 2+ ions was monitored by the decrease in resonance frequency. No adsorption was observed in the absence of Ca 2+ ions, nor even in their presence, if the POPC-POPS mixture was replaced by pure POPC.…”

Section: Alkanethiol-lipid Hybrid Bilayersmentioning

confidence: 99%