3IFD

Human synthetic monocyte chemoattractant protein 1 (MCP-1)

Summary for 3IFD

• Entry DOI: 10.2210/pdb3ifd/pdb
• Descriptor: C-C motif chemokine 2, POTASSIUM ION, PHOSPHATE ION, ... (4 entities in total)
• Functional Keywords: chemokine, quarternary structure, peptide synthesis, chemotaxis, cytokine, disulfide bond, glycoprotein, inflammatory response, pyrrolidone carboxylic acid, secreted
• Biological source: Homo sapiens (human)
• Cellular location: Secreted: P13500
• Total number of polymer chains: 1
• Total formula weight: 8833.11

Authors

Teplyakov, A.,Obmolova, G.,Gilliland, G.L. (deposition date: 2009-07-24, release date: 2009-08-04, Last modification date: 2024-11-06)

Primary citation

Grygiel, T.L.,Teplyakov, A.,Obmolova, G.,Stowell, N.,Holland, R.,Nemeth, J.F.,Pomerantz, S.C.,Kruszynski, M.,Gilliland, G.L.
Synthesis by native chemical ligation and crystal structure of human CCL2.
Biopolymers, 94:350-359, 2010

PubMed Abstract: The protein human CC chemokine ligand 2 (CCL2, also known as monocyte chemoattractant protein 1 or MCP-1) has been synthesized using a combination of solid phase peptide synthesis (SPPS) and native chemical ligation (NCL). The thioester-peptide segment was synthesized using the sulfonamide safety-catch linker and 9-fluorenylmethoxycarbonyl (Fmoc) SPPS, and pseudoproline dipeptides were used to facilitate the synthesis of both CCL2 fragments. After assembly of the full-length peptide chain by NCL, a glutathione redox buffer was used to fold and oxidize the CCL2 protein. Synthetic human CCL2 binds to and activates the CCR2 receptor on THP-1 cells, as expected. CCL2 was crystallized and the structure was determined by X-ray diffraction at 1.9-A resolution. The structure of the synthetic protein is very similar to that of a previously reported structure of recombinant human CCL2, although the crystal form is different. The functional CCL2 dimer for the crystal structure reported here is formed around a crystallographic twofold axis. The dimer interface involves residues Val9-Thr10-Cys11, which form an intersubunit antiparallel beta-sheet. Comparison of the CCL2 dimers in different crystal forms indicates a significant flexibility of the quaternary structure. To our knowledge, this is one of the first crystal structures of a protein prepared using the sulfonamide safety-catch linker and NCL.

PubMed: 20091676
DOI: 10.1002/bip.21390

Experimental method

X-RAY DIFFRACTION (1.9 Å)