Total chemical synthesis and chemotactic activity of human S100A12 (EN-RAGE).
Human S100A12 (extracellular newly identified RAGE (receptor for advanced glycosylation end products)-binding protein), a new member of the S100 family of EF-hand calcium-binding proteins, was chemically synthesised using highly optimised 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate/tert-butoxycarbonyl in situ neutralisation solid-phase chemistry. Circular dichroism studies indicated that CaCl(2) decreased the helical content by 27% whereas helicity was marginally increased by ZnCl(2). The propensity of S100A12 to dimerise was examined by electrospray ionisation time-of-flight mass spectrometry which clearly demonstrated the prevalence of the non-covalent homodimer (20890 Da). Importantly, synthetic human S100A12 in the nanomolar range was chemotactic for neutrophils and macrophages in vitro.
Miranda
L P
LP
Department of Chemistry, Carlsberg Laboratory, Gamle Carlsberg, Valby, Denmark. les@crc.dk
Tao
T
T
Jones
A
A
Chernushevich
I
I
Standing
K G
KG
Geczy
C L
CL
Alewood
P F
PF
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D002630:Chemotactic Factors / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D002633:Chemotaxis / Q000187:drug effects*; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D019281:Dimerization; D018922:HL-60 Cells; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects; D017433:Protein Structure, Secondary / Q000187:drug effects; D009418:S100 Proteins; D000067616:S100A12 Protein; D016415:Sequence Alignment; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology
D002135:Calcium-Binding Proteins; D002630:Chemotactic Factors; D009418:S100 Proteins; D000067616:S100A12 Protein; C090441:S100A12 protein, human; D015032:Zinc; D002118:Calcium