ATP-dependent stabilization and protection of fibroblast growth factor 2

Rose, Karsten; Gast, Ronald E.; Seeger, Anna; Krieglstein, Josef; Klumpp, Susanne

doi:10.1016/j.jbiotec.2009.10.005

Cited by 11 publications

(18 citation statements)

References 31 publications

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“…ATP-binding to FGF2 increased the stability of this growth factor against proteolytic degradation as described previously [12]. Here, we wanted to investigate whether ATP protects the growth factor also from physical inactivation, meaning the stability in aqueous solution.…”

Section: Resultsmentioning

confidence: 83%

“…Recently, we reported that a variant form of FGF2 (FGF2-Quadro, [12]) with mutations in the heparin binding domain did not show enhanced stability against trypsin degradation after incubation with ATP because of its reduced ATP-binding capacity. Here, we investigated the stability of mutant FGF2(K134A) against degradation by neutrophil elastase (NE).…”

Section: Resultsmentioning

confidence: 99%

“…Degradation of FGF2 and FGF2(K134A) by NE was performed as described previously [12]. FGF2 (with his-tag) and FGF2(K134A) (with his-tag) were expressed and purified as described previously [13].…”

Section: Methodsmentioning

confidence: 99%

“…This is similar to the interaction of sulphate residues of heparin or heparan sulphate-proteoglycans (HSPGs) with heparin-binding growth factors like fibroblast growth factors, VEGF, NGF or PDGF [10,11]. Additionally, binding of ATP to FGF2 stabilizes the growth factor and protects it from thermal degradation and proteolytic cleavage with trypsin, neutrophil elastase (NE) or plasmin [12]. …”

Section: Introductionmentioning

confidence: 99%

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Interaction of ATP with fibroblast growth factor 2: biochemical characterization and consequence for growth factor stability

Rose

2011

BMC Biochem

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BackgroundFibroblast growth factor 2, a well-characterized heparin-binding growth factor, is involved in many biological processes like embryogenesis, cell proliferation and angiogenesis. However, this growth factor is very unstable and shows rapid degradation in aqueous solution. Beside the well-known stabilization of FGF2 by heparin or heparan sulphate, the recently discovered binding to ATP also shows a stabilizing and protective effect on this growth factor.ResultsHere we determined the dissociation constant of ATP on FGF2 by equilibrium microdialysis (KD: 59.8 μM) and analyzed the impact of this binding on secondary structure by CD-spectroscopy. ATP-binding to FGF2 significantly changed the secondary structure of this growth factor with a shift to random coil structure elements. We also analyzed the influence of this binding on the stability of FGF2 in aqueous solution over a period of 2 h. While the amount of untreated FGF2 is reduced drastically over this period of time, ATP-binding reduces the degradation considerably.ConclusionsTaken together, our data suggest an important role of ATP in FGF2-stabilization beside the well known-role of heparin and heparan sulphate.

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Section: Resultsmentioning

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Interaction of ATP with fibroblast growth factor 2: biochemical characterization and consequence for growth factor stability

Rose

2011

BMC Biochem

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“…ATP synthase, abundant in the mitochondria, serves as the key enzyme for ATP synthesis. In addition, it plays an important role in protecting fibroblast growth factor 2 (FGF2) from proteolytic cleavage, stabilizing its biologic function in cell proliferation [24,25]. ATP synthase has also been identified on the cell surface of several cell types, including human tumor cells, hepatocytes, keratinocytes, adipocytes, and ECs [26,27].…”

Section: Discussionmentioning

confidence: 99%

The Most Negatively Charged Low-Density Lipoprotein L5 Induces Stress Pathways in Vascular Endothelial Cells

Chen¹,

Hsu²,

Lee

et al. 2012

J Vasc Res

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Background/Aims: L5, the most negatively charged species of low-density lipoprotein (LDL), has been implicated in atherogenesis by inducing apoptosis of endothelial cells (ECs) and inhibiting the differentiation of endothelial progenitor cells. In this study, we compared the effects of LDL charge on cellular stress pathways leading to atherogenesis. Methods: We isolated L5 and L1 (the least negatively charged LDL) from the plasma of patients with familial hypercholesterolemia and used JC-1 staining to examine the effects of L5 and L1 on the mitochondrial membrane potential (DCm) in human umbilical vein ECs (HUVECs). Additionally, we characterized the gene expression profiles of 7 proteins involved in various types of cellular stress. Results: The DCm was severely compromised in HUVECs treated with L5. Furthermore, compared with L1, L5 induced a decrease in mRNA and protein expression of the endoplasmic reticulum (ER) chaperone proteins ORP150, Grp94, and Grp58, mitochondrial proteins Prdx3 and ATP synthase, and an increase in the expression of the pro-inflammatory protein hnRNP C1/C2. Conclusions: Our work suggests that L5, but not L1, may promote the destruction of ECs that occurs during atherogenesis by causing mitochondrial dysfunction and modulating the expression of key proteins to promote inflammation, ER dysfunction, oxidative stress, and apoptosis.

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A Zinc‐Sensing Glucose‐Based Naphthyl Imino Conjugate as a Detecting Agent for Inorganic and Organic Phosphates, Including DNA

Mitra

Hinge

Mittal

et al. 2011

Chemistry A European J

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Owing to the importance of zinc and phosphate in biology and human health, [1] sensing such species is always important and continues to intrigue chemists to emerge with better receptor designs.[2] Our recent report on sensing of Zn 2 + in blood serum milieu by a calixarene derivative [2h] prompted us to employ biologically benign receptor systems and to investigate if these receptors can also extend their sensitivity towards anions (especially phosphates). In this regard, water-soluble and biologically benign [3] carbohydrate-based fluorescent sensors [4] would be of great relevance to explore their in vitro and in vivo applications. Among the derivatives of phosphate, adenosine-mono/di/triphosphate (AMP, ADP, ATP), DNA, and their hydrolytic species are important.[5] Therefore, it is essential to monitor the presence of phosphate related species in solution.[6] If a receptor can sense both the zinc and the phosphate group, it would be doubly advantageous.[5e] Therefore, herein we investigate the primary detection of Zn 2 + by switch-on fluorescence using 1-(d-glucopyranosyl-2'-deoxy-2'-iminomethyl)-2-hydroxynaphthalene conjugate L and the successive detection of phosphates by the corresponding zinc complex through fluorescence quenching (switch-off). To demonstrate the bio-suitability of L, titrations were carried out using nucleotides and DNA. To our knowledge, this is the first carbohydrate-based receptor system shown to be sensitive towards a cation followed by an anion, including biomolecular systems.Conjugate L has been synthesized by a one-pot condensation of the neutralized C2-deoxy-C2-amino-glucose with 2-hydroxy-1-naphthaldehyde and his structure has been characterized (see SI 01 in the Supporting Information).[7] Conjugate L has been crystallized in a non-centrosymmetric triclinic system by layering methanol on to a concentrated solution in DMSO and the structure was determined by single crystal XRD (for crystal and metric data see SI 02 in the Supporting Information). Conjugate L exhibits a preformed ONO core that is suitable for metal ion binding (Scheme 1). An intramolecular hydrogen bond present between the phenolic O4 À H···N1-imino orients the naphthyl moiety almost perpendicular to the mean plane of the carbohydrate moiety (see SI 02b in the Supporting Information). In the lattice, molecules form columns by interacting through two strong hydrogen bonds per each L and also by exhibiting hydrophobic p-p interactions with a centroid to centroid distance of 4.878 . The columnar structure formed in the lattice clearly exhibits hydrophobic (6 ) and hydrophilic (7 ) regions alternatively (see SI 02c in the Supporting Information).The selectivity of L has been demonstrated by titration against seven biologically and ecologically relevant metal ions, such as, Na + , K + , Ca 2 + , Mg 2 + , Zn 2 + , Cd 2 + , and Hg 2 + in methanol using fluorescence spectroscopy. In dry methanol, L exhibits approximately a 140-fold enhancement in the emission intensity against Zn 2 + , while none of the alkali and al...

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ATP-dependent stabilization and protection of fibroblast growth factor 2

Cited by 11 publications

References 31 publications

Interaction of ATP with fibroblast growth factor 2: biochemical characterization and consequence for growth factor stability

Interaction of ATP with fibroblast growth factor 2: biochemical characterization and consequence for growth factor stability

The Most Negatively Charged Low-Density Lipoprotein L5 Induces Stress Pathways in Vascular Endothelial Cells

A Zinc‐Sensing Glucose‐Based Naphthyl Imino Conjugate as a Detecting Agent for Inorganic and Organic Phosphates, Including DNA

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