Expression, characterization, and preliminary X-ray crystallographic analysis of recombinant murine Follistatin-like 1 expressed in Drosophila S2 cells

Li, Lian; Li, Xinxin; Xue, Lixiang; Dong, Yingying; Geng, Yan; Liu, Xinqi; Ning, Wen

doi:10.5582/bst.2013.v7.2.93

Cited by 4 publications

(6 citation statements)

References 35 publications

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“…The full‐length Fstl1 without leading peptide was expressed in Drosophila S2 cells in secreted form, and purified protein was used for crystallization. The crystals appeared in crystallization drops were used for data collection and structure determination . Unfortunately, after solving the structure, we found that only N‐terminal FOLN‐Kazal region were present in the crystal structure (Figure a), so we analyzed this structure and studied the function of this domain subsequently.…”

Section: Resultssupporting

confidence: 81%

“…Co‐immunoprecipitate of GFP‐tagged and HA‐tagged Fstl1‐FK also agrees with the existence of Fstl1 dimer in living cells (Figure e). Consistent with this observation, we have previously reported that purified full‐length Fstl1 also forms dimer in solution . The FK domain from Follistatin can also be eluted as both monomer and dimer in gel filtration column, indicating these protein might play their roles in a dimer form.…”

Section: Resultsmentioning

confidence: 99%

“…The crystals appeared in crystallization drops were used for data collection and structure determination. 30 Unfortunately, after solving the structure, we found that only N-terminal FOLN-Kazal region were present in the crystal structure (Figure 1a), so we analyzed this structure and studied the function of this domain subsequently.…”

Section: Crystal Structure Of Fstl1-fk Domainmentioning

confidence: 99%

“…Consistent with this observation, we have previously reported that purified full-length Fstl1 also forms dimer in solution. 30 The FK (e) Co-immunoprecipitate of GFP-Fstl1-FK and HA-Fstl1-FK. Two proteins were co-expressed in 293T cells and pulled-down by GFP-trap beads, then detected by anti-HA antibody domain from Follistatin can also be eluted as both monomer and dimer in gel filtration column, indicating these protein might play their roles in a dimer form.…”

Section: Dimerization Of Fstl1-fkmentioning

confidence: 99%

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Structural and functional study of FK domain of Fstl1

Chang

et al. 2019

Protein Science

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Fstl1 is a TGF‐β superfamily binding protein which involved in many pathological processes. The function of Fstl1 has been widely elucidated, but its structural characterization has not been explored. Here we solved the high‐resolution crystal structure of FK domain of murine Fstl1, analyzed its unique characteristics, and investigated its contribution to the function of full‐length Fstl1. We found that Fstl1‐FK forms a stable dimer in both solution and crystal, which suggest that this protein may function as a dimer during its interaction with TGF‐β, a molecule known to form dimer during activation process. We also found this FK domain is indispensable for the proper function of Fstl1 during the transduction of TGF‐β signaling. These observations provide important insights into the understanding of Fstl1 and may facilitate the exploration of this molecule in clinical study.

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Section: Resultssupporting

confidence: 81%

Section: Resultsmentioning

confidence: 99%

Section: Crystal Structure Of Fstl1-fk Domainmentioning

confidence: 99%

Section: Dimerization Of Fstl1-fkmentioning

confidence: 99%

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Structural and functional study of FK domain of Fstl1

Chang

et al. 2019

Protein Science

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“…Cells of passages 3–4 were used for myofibroblast activation and ECM production assays. After the indicated length of time of 5 ng/ml TGF-β1 treatment with/without 100 ng/ml FSTL1 protein ( Li et al, 2013 ), fibroblasts and cultured medium were collected separately, and the α-SMA expression in cell extracts or the ECM production in supernatants of medium was measured using Western blot analysis.…”

Section: Methodsmentioning

confidence: 99%