Expression and crystallographic studies of the D1D2 domains of C4.4A, a homologous protein to the urokinase receptor

TY - JOUR

T1 - Expression and crystallographic studies of the D1D2 domains of C4.4A, a homologous protein to the urokinase receptor

AU - Chen, Shanli

AU - Lin, Lin

AU - Yuan, Cai

AU - Gärdsvoll, Henrik

AU - Kriegbaum, Mette C.

AU - Ploug, Michael

AU - Huang, Mingdong

PY - 2017/8/1

Y1 - 2017/8/1

N2 - C4.4A is a glycosylphosphatidylinositol-anchored membrane protein comprised of two LU domains (Ly6/uPAR-like domains) and an extensively O-glycosylated C-terminal Ser/Thr/Pro-rich region. C4.4A is a novel biomarker for squamous epithelial differentiation. Its expression is dysregulated under various pathological conditions and it is a robust biomarker for poor prognosis in various malignant conditions such as pulmonary adenocarcinoma. To facilitate crystallization, the two LU domains were excised from intact C4.4A by limited proteolysis, purified and crystallized by the sitting-drop vapour-diffusion method. The crystals diffracted to 2.7 Å resolution and belonged to space group C2221, with unit-cell parameters a = 55.49, b = 119.63, c = 168.54 Å. The statistics indicated good quality of the data, which form a solid basis for the determination of the C4.4A structure.The two amino-terminal LU domains of C4.4A were excised by limited proteolysis and were crystallized using the sitting-drop vapour-diffusion method. The crystals diffracted to 2.7 Å resolution, giving good-quality data.

AB - C4.4A is a glycosylphosphatidylinositol-anchored membrane protein comprised of two LU domains (Ly6/uPAR-like domains) and an extensively O-glycosylated C-terminal Ser/Thr/Pro-rich region. C4.4A is a novel biomarker for squamous epithelial differentiation. Its expression is dysregulated under various pathological conditions and it is a robust biomarker for poor prognosis in various malignant conditions such as pulmonary adenocarcinoma. To facilitate crystallization, the two LU domains were excised from intact C4.4A by limited proteolysis, purified and crystallized by the sitting-drop vapour-diffusion method. The crystals diffracted to 2.7 Å resolution and belonged to space group C2221, with unit-cell parameters a = 55.49, b = 119.63, c = 168.54 Å. The statistics indicated good quality of the data, which form a solid basis for the determination of the C4.4A structure.The two amino-terminal LU domains of C4.4A were excised by limited proteolysis and were crystallized using the sitting-drop vapour-diffusion method. The crystals diffracted to 2.7 Å resolution, giving good-quality data.

KW - C4.4A

KW - glycosylation

KW - LU domains

KW - prognostic cancer biomarkers

U2 - 10.1107/S2053230X17009748

DO - 10.1107/S2053230X17009748

M3 - Journal article

C2 - 28777093

AN - SCOPUS:85026822472

SN - 2053-230X

VL - 73

SP - 486

EP - 490

JO - Acta Crystallographica Section F: Structural Biology Communications

JF - Acta Crystallographica Section F: Structural Biology Communications

IS - 8

ER -