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

Shanli Chen; Lin Lin; Cai Yuan; Henrik Gårdsvoll; Mette C Kriegbaum; Michael Ploug; Mingdong Huang

doi:10.1107/S2053230X17009748

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

Acta Crystallogr F Struct Biol Commun. 2017 Aug 1;73(Pt 8):486-490. doi: 10.1107/S2053230X17009748. Epub 2017 Jul 27.

Authors

Shanli Chen¹, Lin Lin², Cai Yuan³, Henrik Gårdsvoll⁴, Mette C Kriegbaum⁴, Michael Ploug⁴, Mingdong Huang¹

Affiliations

¹ College of Chemistry, Fuzhou Univerisity, Fuzhou, Fujian 350108, People's Republic of China.
² Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02115, USA.
³ College of Bioscience and Biotechnology, Fuzhou Univerisity, Fuzhou, Fujian 350108, People's Republic of China.
⁴ The Finsen Laboratory, Rigshospitalet, DK-2200 Copenhagen, Denmark.

Abstract

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.

Keywords: C4.4A; LU domains; glycosylation; prognostic cancer biomarkers.

MeSH terms

Amino Acid Sequence*
Animals
Biomarkers, Tumor / chemistry*
Biomarkers, Tumor / genetics
Biomarkers, Tumor / metabolism
Cell Adhesion Molecules / chemistry*
Cell Adhesion Molecules / genetics
Cell Adhesion Molecules / metabolism
Cloning, Molecular
Crystallization
Crystallography, X-Ray
Drosophila melanogaster / cytology
Drosophila melanogaster / metabolism
GPI-Linked Proteins / chemistry
GPI-Linked Proteins / genetics
GPI-Linked Proteins / metabolism
Gene Expression
Genetic Vectors / chemistry
Genetic Vectors / metabolism
Humans
Protein Domains
Receptors, Urokinase Plasminogen Activator / chemistry
Receptors, Urokinase Plasminogen Activator / genetics
Receptors, Urokinase Plasminogen Activator / metabolism
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Sequence Deletion*
Sequence Homology, Amino Acid
X-Ray Diffraction

Substances

Biomarkers, Tumor
Cell Adhesion Molecules
GPI-Linked Proteins
LYPD3 protein, human
Receptors, Urokinase Plasminogen Activator
Recombinant Proteins

Abstract

MeSH terms

Substances

Grants and funding