Physical Stabilization of Acidic Fibroblast Growth Factor by Polyanions

Volkin, David B.; Tsai, Pei-Kuo; Dabora, Jonathan M.; Gress, Jacqueline; Burke, Catherine; Linhardt, Robert J.; Middaugh, C. Russell

doi:10.1006/abbi.1993.1005

Cited by 121 publications

(126 citation statements)

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“…This electrostatic description of the role of heparin is entirely consistent with the early report by Linhardt and Middaugh that the thermal stability of aFGF was enhanced by not only heparin but also "a surprising number of polyanions." 63 If the aggregation of insulin is subject to equilibrium, as indicated by the lack of significant hysteresis upon back-titration with NaOH for the system in Figure 1 (see Supporting Information), the binding of the insulin dimer to heparin should shift the equilibrium away from aggregate. Thus, heparin can reverse the aggregation of insulin, by the mechanism illustrated in Figure 5B.…”

Section: Discussionmentioning

confidence: 99%

Suppression of Insulin Aggregation by Heparin

et al. 2008

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The aggregation of insulin near its isoelectric point and at low ionic strength was suppressed in the presence of heparin. To understand this effect, we used turbidimetry and stopped-flow to study the pH-and ionic strength (I)-dependence of the aggregation of heparin-free insulin. The results supported the role of interprotein electrostatic interactions, contrary to the commonly held view that such forces are minimized at pH ) pI. Electrostatic modeling of insulin (DelPhi) revealed that attractive interactions arise from the marked charge anisotropy of insulin near pI. We show how screening of the interprotein attractions by added salt lead to maximum aggregation near I ) 0.01 M, corresponding to a Debye length nearly equal to the diameter of the insulin dimer, consistent with a dipole-like protein charge distribution. This analysis is also consistent with suppression of aggregation by heparin, a strong polyanion that by binding to the positive domain of one protein, inhibits its interaction with the negative domain of another.

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Section: Discussionmentioning

confidence: 99%

Suppression of Insulin Aggregation by Heparin

et al. 2008

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“…Thus, a rational therapeutic strategy to inhibit amyloidosis is to design ligands that bind with high affinity and specificity to the native protein (32,33). Relative to the effects of 1.0 M sucrose, stoichiometric amounts these compounds greatly increase the thermodynamic stability of the native state and reduce the levels of partially and fully unfolded protein molecules (15,32,33,36,37). In this regard, naturally occurring stabilizing ligands have been shown to inhibit amyloid fibril formation by transthyretin (32,33), as well as, for example, the formation of insoluble aggregates by acid-fibroblast growth factor (36).…”

Section: Discussionmentioning

confidence: 99%

“…Relative to the effects of 1.0 M sucrose, stoichiometric amounts these compounds greatly increase the thermodynamic stability of the native state and reduce the levels of partially and fully unfolded protein molecules (15,32,33,36,37). In this regard, naturally occurring stabilizing ligands have been shown to inhibit amyloid fibril formation by transthyretin (32,33), as well as, for example, the formation of insoluble aggregates by acid-fibroblast growth factor (36). For most amyloidogenic proteins, high affinity ligands have not been identified but can be designed using combinatorial library screening and protein structure-based methods (e.g.…”

Section: Discussionmentioning

confidence: 99%

Thermodynamic Modulation of Light Chain Amyloid Fibril Formation

Kim¹,

Wall²,

Meyer³

et al. 2000

Journal of Biological Chemistry

133

131

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To obtain further insight into the pathogenesis of amyloidosis and develop therapeutic strategies to inhibit fibril formation we investigated: 1) the relationship between intrinsic physical properties (thermodynamic stability and hydrogen-deuterium (H-D) exchange rates) and the propensity of human immunoglobulin light chains to form amyloid fibrils in vitro; and 2) the effects of extrinsically modulating these properties on fibril formation. An amyloid-associated protein readily formed amyloid fibrils in vitro and had a lower free energy of unfolding than a homologous nonpathological protein, which did not form fibrils in vitro. H-D exchange was much faster for the pathological protein, suggesting it had a greater fraction of partially folded molecules. The thermodynamic stabilizer sucrose completely inhibited fibril formation by the pathological protein and shifted the values for its physical parameters to those measured for the nonpathological protein in buffer alone. Conversely, urea sufficiently destabilized the nonpathological protein such that its measured physical properties were equivalent to those of the pathological protein in buffer, and it formed fibrils. Thus, fibril formation by light chains is predominantly controlled by thermodynamic stability; and a rational strategy to inhibit amyloidosis is to design high affinity ligands that specifically increase the stability of the native protein.

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“…To experimentally test this, we generated an oxidized Sp100 PB by Cu 2ϩ -catalyzed disulfide bond formation (24). To avoid unwanted intermolecular cross-linking, we used a surface triple mutant Sp100C PB (C754S/L772Q/C793S, originally introduced to facilitate crystallization), in which two surface cysteine residues were mutated to serine.…”

Section: Mutagenesis and H3k4me0 Co-localization Analysis-mentioning

confidence: 99%

Multifaceted Histone H3 Methylation and Phosphorylation Readout by the Plant Homeodomain Finger of Human Nuclear Antigen Sp100C

Zhang

Zhao

Xiong

et al. 2016

Journal of Biological Chemistry

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The decoding of histone post-translational modifications by chromatin-binding modules ("readers") constitutes one major mechanism of epigenetic regulation. Nuclear antigen Sp100 (SPECKLED, 100 kDa), a constitutive component of the promyelocytic leukemia nuclear bodies, plays key roles in intrinsic immunity and transcriptional repression. Sp100C, a splicing isoform specifically up-regulated upon interferon stimulation, harbors a unique tandem plant homeodomain (PHD) finger and bromodomain at its C terminus. Combining structural, quantitative binding, and cellular co-localization studies, we characterized Sp100C PHD finger as an unmethylated histone H3 Lys 4 (H3K4me0) reader that tolerates histone H3 Thr 3 phosphorylation (H3T3ph), histone H3 Lys 9 trimethylation (H3K9me3), and histone H3 Ser 10 phosphorylation (H3S10ph), hallmarks associated with the mitotic chromosome. In contrast, whereas H3K4me0 reader activity is conserved in Sp140, an Sp100C paralog, the multivalent tolerance of H3T3ph, H3K9me3, and H3S10ph was lost for Sp140. The complex structure determined at 2.1 Å revealed a highly coordinated lysine ⑀-amine recognition sphere formed by an extended N-terminal motif for H3K4me0 readout. Interestingly, reader pocket rigidification by disulfide bond formation enhanced H3K4me0 binding by Sp100C. An additional complex structure solved at 2.7 Å revealed that H3T3ph is recognized by the arginine residue, Arg 713 , that is unique to the PHD finger of Sp100C. Consistent with a restrictive cellular role of Sp100C, these results establish a direct chromatin targeting function of Sp100C that may regulate transcriptional gene silencing and promyelocytic leukemia nuclear body-mediated intrinsic immunity in response to interferon stimulation.

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Physical Stabilization of Acidic Fibroblast Growth Factor by Polyanions

Cited by 121 publications

References 0 publications

Suppression of Insulin Aggregation by Heparin

Suppression of Insulin Aggregation by Heparin

Thermodynamic Modulation of Light Chain Amyloid Fibril Formation

Multifaceted Histone H3 Methylation and Phosphorylation Readout by the Plant Homeodomain Finger of Human Nuclear Antigen Sp100C

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