Crystallization and preliminary X-ray crystallographic analysis of SEDL.
SEDL (known also as sedlin) is a 140 amino-acid protein with a putative role in endoplasmic reticulum-to-Golgi transport. Several missense mutations and deletion mutations in the SEDL gene, which result in protein truncation by frame shift, are responsible for spondyloepiphyseal dysplasia tarda, a progressive skeletal disorder. The protein is identical to MIP-2A, which was shown to interact physically with c-myc promotor-binding protein 1 (MBP-1) and relieve the regulatory role of MBP-1 as a general transcription repressor. In order to gain insights into the function of SEDL by structural analysis, the protein was overexpressed and crystallized as a first step. SEDL was overexpressed in Escherichia coli and crystallized using the hanging-drop vapour-diffusion method at 298 K. The crystals belong to the orthorhombic space group C222(1), with unit-cell parameters a = 46.69, b = 101.30, c = 66.15 A. The unit cell is likely to contain one molecule of SEDL, with a crystal volume per protein mass (V(M)) of 2.36 A(3)Da(-1) and a solvent content of about 47.9% by volume. A native data set to 2.8 A resolution was obtained from a flash-cooled crystal using synchrotron radiation.
Jang
Se Bok
SB
National Creative Research Initiative Center for Biomolecular Recognition and Department of Life Science, Pohang University of Science and Technology, Pohang, Kyungbuk 790-784, South Korea.
Cho
Yong-Soon
YS
Eom
Soo-Jung
SJ
Choi
Eui-Ju
EJ
Kim
Kyung-Hwa
KH
Suh
Pann-Ghill
PG
Oh
Byung-Ha
BH
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D026901:Membrane Transport Proteins; D051379:Mice; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D013045:Species Specificity; D014157:Transcription Factors
D002352:Carrier Proteins; D026901:Membrane Transport Proteins; D011994:Recombinant Proteins; C120759:TRAPPC2 protein, human; D014157:Transcription Factors