Crystal structure of recombinant human stromal cell‐derived factor‐1α

Ryu, Eui Kyung; Kim, Tae Kyun; Kwon, Tai Hun; Jung, In Duk; Ryu, Dowook; Park, Yeong‐Min; Kim, Junhong; Ahn, Kyo Han; Ban, Changill

doi:10.1002/prot.21350

Cited by 28 publications

(33 citation statements)

References 20 publications

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“…1(A)]. This configuration is incompatible with the CXC-type dimer observed in subsequent SDF-1 crystal structures 25,30,31 [ Fig. 1 (helix angle ¼ 90 ) and others that are not (helix angle < 90 ).…”

Section: Introductionmentioning

confidence: 72%

“…Cyanogen bromide cleavage to generate biologically active SDF-1 27,28,45 was omitted for these structural studies because the flexible SDF-1 N-terminus does not interact with the folded chemokine domain or influence the monomer-dimer equilibrium. [24][25][26][27]32 Hence, SDF-1 used for structural studies contained an N-terminal Gly-Ser dipeptide for the structure solved in 25 mM MES at pH 6.8 or an N-terminal gly-met dipeptide for the structures solved in either 0 mM sodium phosphate at pH 5.5 or 50 mM sodium phosphate at pH 5.5.…”

Section: Methodsmentioning

confidence: 99%

“…12 In each case, SDF-1 treatment is effective, but variations in the protective effect may be dose-related. SDF-1 exists in a monomer-dimer equilibrium that shifts toward dimer upon crystallization [23][24][25] and in the presence of various binding partners, 26 including an extracellular domain of its receptor CXCR4. 27,28 To investigate the structure-function relationships of the two oligomeric states, we generated a symmetric disulfide-linked dimer [SDF-1(L36C/A65C)] 27 and a preferentially monomeric variant (H25R).…”

Section: Introductionmentioning

confidence: 99%

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Monomeric structure of the cardioprotective chemokine SDF‐1/CXCL12

Veldkamp

Ziarek

et al. 2009

Protein Science

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The chemokine stromal cell-derived factor-1 (SDF-1/CXCL12) directs leukocyte migration, stem cell homing, and cancer metastasis through activation of CXCR4, which is also a coreceptor for T-tropic HIV-1. Recently, SDF-1 was shown to play a protective role after myocardial infarction, and the protein is a candidate for development of new anti-ischemic compounds. SDF-1 is monomeric at nanomolar concentrations but binding partners promote self-association at higher concentrations to form a typical CXC chemokine homodimer. Two NMR structures have been reported for the SDF-1 monomer, but only one matches the conformation observed in a series of dimeric crystal structures. In the other model, the C-terminal helix is tilted at an angle incompatible with SDF-1 dimerization. Using a rat heart explant model for ischemia/ reperfusion injury, we found that dimeric SDF-1 exerts no cardioprotective effect, suggesting that the active species is monomeric. To resolve the discrepancy between existing models, we solved the NMR structure of the SDF-1 monomer in different solution conditions. Irrespective of pH and buffer composition, the C-terminal helix remains tilted at an angle with no evidence for the perpendicular arrangement. Furthermore, we find that phospholipid bicelles promote dimerization that necessarily shifts the helix to the perpendicular orientation, yielding dipolar couplings that are incompatible with the NOE distance constraints. We conclude that interactions with the alignment medium biased the previous structure, masking flexibility in the helix position that may be essential for the distinct functional properties of the SDF-1 monomer.

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Section: Introductionmentioning

confidence: 72%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Monomeric structure of the cardioprotective chemokine SDF‐1/CXCL12

Veldkamp

Ziarek

et al. 2009

Protein Science

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“…vCCI) (Carfí et al, 1999). Structural modeling was carried out using the following PDB entries: PDB: 2NYZ (XCL1) (Alexander-Brett and Fremont, 2007), PDB: 1EL0 (CCL1) (Keizer et al, 2000), PDB: 1QNK (CXCL2) (Qian et al, 1999), PDB: 1RHP (CXCL4) (Zhang et al, 1994), PDB: 2IL8 (CXCL8) (Clore et al, 1990), PDB: 1O7Z (CXCL10) (Swaminathan et al, 2003), and PDB: 2J7Z (CXCL12) (Ryu et al, 2007). The structural models of ORFV CKBP bound to various chemokines were based on the 2.60-Å ORFV CKBP-CCL2 structure using simple rigid body superpositions.…”

Section: Structural Analysis and Modelingmentioning

confidence: 99%

Structures of Orf Virus Chemokine Binding Protein in Complex with Host Chemokines Reveal Clues to Broad Binding Specificity

et al. 2015

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The chemokine binding protein (CKBP) from orf virus (ORFV) binds with high affinity to chemokines from three classes, C, CC, and CXC, making it unique among poxvirus CKBPs described to date. We present its crystal structure alone and in complex with three CC chemokines, CCL2, CCL3, and CCL7. ORFV CKBP possesses a β-sandwich fold that is electrostatically and sterically complementary to its binding partners. Chemokines bind primarily through interactions involving the N-terminal loop and a hydrophobic recess on the ORFV CKBP β-sheet II surface, and largely polar interactions between the chemokine 20s loop and a negatively charged surface groove located at one end of the CKBP β-sheet II surface. ORFV CKBP interacts with leukocyte receptor and glycosaminoglycan binding sites found on the surface of bound chemokines. SEC-MALLS and chromatographic evidence is presented supporting that ORFV CKBP is a dimer in solution over a broad range of protein concentrations.

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“…Only 2 of these variants of SDF-1 showed significant activity on cell migration, a finding that can be explained by the fact that the receptor binding side of SDF-1 is located at the N-terminus. 16 We performed in vitro cell migration assays and in vivo Matrigel plug assays that indicated that SSDF-1(S4V) was the most potent mutant of SDF-1 identified with this strategy. We also performed receptor activation assays with SSDF-1(S4V) and showed that mutating the serine at position 4 to a valine and addition of an N-terminal serine yields a protein that is an agonist of both CXCR4 and CXCR7 receptors, although with a slightly lower potency on CXCR4 than native SDF-1.…”

Section: Discussionmentioning

confidence: 99%

Protease-Resistant Stromal Cell–Derived Factor-1 for the Treatment of Experimental Peripheral Artery Disease

Segers

Revin

et al. 2011

Circulation

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Background-Peripheral artery disease is a potentially incapacitating disease for which pharmacological options are limited.Stromal cell-derived factor-1 (SDF-1) is a chemokine that attracts endothelial progenitor cells and promotes angiogenesis. Therapeutic use of SDF-1 in hindlimb ischemia may be challenged by proteolytic degradation. We hypothesized that protease-resistant variants of SDF-1 can increase blood flow in an experimental model of hindlimb ischemia. Methods and Results-We screened a peptide library for mutations in SDF-1 that provide resistance to matrix metalloproteinase cleavage. Recombinant SDF-1 proteins carrying the mutations were designed, expressed, and purified, and activity of mutant proteins was tested with receptor activation assays and in vivo Matrigel plug assays. SSDF-1(S4V), which is resistant to both dipeptidylpeptidase IV/CD26 and matrix metalloproteinase-2 cleavage, was active in vitro and induced angiogenesis in vivo. We then designed and purified fusion proteins of SSDF-1 and SSDF-1(S4V) with the sequence of self-assembling peptide nanofibers for incorporation into nanofibers. In a blinded and randomized hindlimb ischemia mouse study, SSDF-1(S4V) delivery by nanofibers improved blood flow as measured by laser Doppler from 23.1Ϯ1.9% (untreated control) to 55.1Ϯ5.7% 6 weeks after surgery (PϽ0.001). Nanofibers alone or SSDF-1 delivered by nanofibers did not improve blood flow. Furthermore, SSDF-1(S4V) delivered by nanofibers increased formation of new arterioles. In vitro, SSDF-1(S4V) attracts smooth muscle cells but does not induce mitosis. Conclusions-SDF-1 engineered to be resistant to dipeptidylpeptidase IV/CD26 and matrix metalloproteinase-2 cleavage and delivered by nanofibers improves blood flow in a model of peripheral artery disease. (Circulation. 2011;123:1306-1315.)

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Crystal structure of recombinant human stromal cell‐derived factor‐1α

Cited by 28 publications

References 20 publications

Monomeric structure of the cardioprotective chemokine SDF‐1/CXCL12

Monomeric structure of the cardioprotective chemokine SDF‐1/CXCL12

Structures of Orf Virus Chemokine Binding Protein in Complex with Host Chemokines Reveal Clues to Broad Binding Specificity

Protease-Resistant Stromal Cell–Derived Factor-1 for the Treatment of Experimental Peripheral Artery Disease

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