Synthesis by native chemical ligation and crystal structure of human CCL2.
Biopolymers
; 94(3): 350-9, 2010.
Article
in En
| MEDLINE
| ID: mdl-20091676
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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Protein Structure, Tertiary
/
Chemokine CCL2
/
Protein Structure, Quaternary
Type of study:
Prognostic_studies
Limits:
Humans
Language:
En
Journal:
Biopolymers
Year:
2010
Document type:
Article
Affiliation country:
Estados Unidos