Oligomeric self-association of basic fibroblast growth factor in the absence of heparin-like glycosaminoglycans

Davis, Joseph; Venkataraman, Ganesh; Shriver, Zachary; Raj, Periathamby Antony; Sasisekharan, Ram

doi:10.1042/0264-6021:3410613

Cited by 13 publications

(15 citation statements)

References 33 publications

(41 reference statements)

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“…According to this structural model, the binding of heparin to FGF7 would cause a folding of the molecule allowing KFGR recognition. However, depending on the cell context, HS might not be required for binding and internalization of other FGFs, such as FGF2 [53], and FGF2 oligomers can be formed in the absence of HS [54]. Moreover, FGF2-FGFR crystals can form in the absence of heparin [55].…”

Section: Discussionmentioning

confidence: 99%

A role for the perlecan protein core in the activation of the keratinocyte growth factor receptor

2001

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Perlecan, a widespread heparan sulphate (HS) proteoglycan, is directly involved in the storing of angiogenic growth factors, mostly members of the fibroblast growth factor (FGF) gene family. We have previously shown that antisense targeting of the perlecan gene causes a reduced growth and responsiveness to FGF7 [also known as keratinocyte growth factor (KGF)] in human cancer cells, and that the perlecan protein core interacts specifically with FGF7. In the present paper, we have investigated human colon carcinoma cells in which the perlecan gene was disrupted by targeted homologous recombination. After screening over 1000 clones, we obtained two clones heterozygous for the null mutation with no detectable perlecan, indicating that the other allele was non-functioning. The perlecan-deficient cells grew more slowly, did not respond to FGF7 with or without the addition of heparin, and were less tumorigenic than control cells. Paradoxically, the perlecan-deficient cells displayed increased FGF7 surface binding. However, the perlecan protein core was required for functional activation of the KGF receptor and downstream signalling. Because heparin could not substitute for perlecan, the HS chains are not critical for FGF7-mediated signalling in this cell system. These results provide the first genetic evidence that the perlecan protein core is a molecular entity implicated in FGF7 binding and activation of its receptor.

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Section: Discussionmentioning

confidence: 99%

A role for the perlecan protein core in the activation of the keratinocyte growth factor receptor

2001

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“…In addition, HLGAGs are involved in facilitating FGF2 binding and activation (37,38). We and others have previously demonstrated that HLGAGs serve to induce dimerization and oligomerization of FGF2 (27,(39)(40)(41), presumably to facilitate receptor dimerization. In addition, HLGAGs are involved in the internalization of FGF2 for intracellular processing (42,43) and in the control of FGF2 transport within the ECM (44).…”

Section: Roles Of Hlgags In Modulating Fgf2 Activitymentioning

confidence: 94%

Distinct heparan sulfate glycosaminoglycans are responsible for mediating fibroblast growth factor‐2 biological activity through different fibroblast growth factor receptors

et al. 2001

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Fibroblast growth factor (FGF) signaling is involved in many important biological processes such as smooth muscle cell (SMC) proliferation, a key event leading to atherosclerosis. It is well known that heparin/heparan sulfate‐like glycosaminoglycans (HLGAGs), found ubiquitously on the cell surface and in the extracellular matrix, modulate FGF signaling by interacting with both FGF and FGF receptor (FGFR). To study the modulatory effect of SMC‐derived HLGAGs on FGF2 signaling, we employed a panel of HLGAG‐degrading enzymes, or heparinases, to depolymerize cell surface HLGAGs isolated from SMCs and assayed their abilities to modulate FGF2 activity on engineered cell lines expressing defined FGFR isoforms. Our findings indicate that FGF2‐mediated proliferation is dependent on the composition of the SMC‐derived HLGAG fragments as well as the type of FGFR isoform expressed. We postulate that the chemical composition and sequence of HLGAGs dictate the specificity of interactions between FGF2 and a given FGFR.

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“…Glutaraldehyde Crosslinking reaction-Crosslinking reaction was performed to stabilize the oligomer form of the protein which can be distinguished from the monomer form following SDS-PAGE analysis. Glutaraldehyde was chosen for chemical cross-linking for reasons that included aqueous solubility, homobifunctional specificity for lysine residues, short spacer length (5Å) (30). A typical reaction contained 5µg (0.33 µmoles)…”

Section: Downloaded Frommentioning

confidence: 99%

Expression and molecular characterization of the Mycobacterium tuberculosis PII protein

Bandyopadhyay

Arora

Jain

et al. 2010

The Journal of Biochemistry

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The signal transduction protein PII plays an important role in cellular nitrogen assimilation and regulation. The molecular characteristics of the Mycobacterium tuberculosis PII (Mtb PII) were investigated using biophysical experiments. The Mtb PII coding ORF Rv2919c was cloned and expressed in Escherichia coli. The binding characteristics of the purified protein with ATP and ADP were investigated using surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC). Mtb PII binds to ATP strongly with K(d) in the range 1.93-6.44 microM. This binding strength was not significantly affected by the presence of 2-ketoglutarate even in molar concentrations of 66 (ITC) or 636 (SPR) fold excess of protein concentration. However, an additional enthalpy of 0.3 kcal/mol was released in presence of 2-ketoglutarate. Binding of Mtb PII to ADP was weaker by an order of magnitude. Binding of ATP and 2-ketoglutarate were analysed by docking studies on the Mtb PII crystal structure (PDB id 3BZQ). We observed that hydrogen bonds involving the gamma-phosphate of ATP contribute to enhanced binding of ATP compared with ADP. Glutaraldehyde crosslinking showed that Mtb PII exists in homotrimeric state which is consistent with other PII proteins. Phylogenetic analysis showed that Mtb PII consistently grouped with other actinobacterial PII proteins.

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Oligomeric self-association of basic fibroblast growth factor in the absence of heparin-like glycosaminoglycans

Cited by 13 publications

References 33 publications

A role for the perlecan protein core in the activation of the keratinocyte growth factor receptor

A role for the perlecan protein core in the activation of the keratinocyte growth factor receptor

Distinct heparan sulfate glycosaminoglycans are responsible for mediating fibroblast growth factor‐2 biological activity through different fibroblast growth factor receptors

Expression and molecular characterization of the Mycobacterium tuberculosis PII protein

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