TY - JOUR
T1 - Synthesis and characterization of indolicidin, a tryptophan‐rich antimicrobial peptide from bovine neutrophils
AU - ABEL, ROBERT J.VAN
AU - TANG, YI‐QUAN ‐Q
AU - RAO, V. S.V.
AU - DOBBS, CRAIG H.
AU - TRAN, DAT
AU - Barany, George
AU - SELSTED, MICHAEL E.
PY - 1995/5
Y1 - 1995/5
N2 - Indolicidin, a novel tryptophan‐rich microbicidal tridecapeptide amide isolated originally from granules of bovine neutrophils, has been prepared by optimized manual and automated protocols of stepwise solid‐phase synthesis with Nα‐9‐fluorenylmethyloxycarbonyl (Fmoc) amino acid derivatives. Both standard polystyrene (PS) and polyethylene glycol‐polystyrene (PEG‐PS) graft supports were used in combination with handles that provide C‐terminal peptide amides: 5‐(4‐Fmoc‐aminomethyl‐3,5‐dimethoxyphenoxy)valeric acid (PAL) or 5‐(9‐Fmoc‐aminoxanthen‐2‐oxy)valeric acid (XAL). Final deprotection/cleavage was carried out with reagent K, trifluoroacetic acid–phenol–water–thioanisole–1,2‐ethanedithiol (82.5:5:5:5:2.5), or reagent B, trifluoroacetic acid–phenol–water–tri(isopropyl)silane (88:5:5:2), and related cocktails. Initial purities as high as 93% were obtained immediately following cleavage. In the largest‐scale synthesis carried out, 0.8 g of HPLC‐purified indolicidin (> 99% pure) was obtained, representing a 39% overall yield based on C‐terminal Arg(Pmc) anchored to PAL‐PS‐resin. The main synthetic product, and some by‐products, were characterized by analytical high‐performance liquid chromatography (HPLC), sequencing, and fast atom bombardment mass spectrometry (FABMS). The antimicrobial potencies of natural and synthetic indolicidin, as determined by in vitro antibacterial and antifungal assays, were identical. Further, the reactivities of natural and synthetic peptides with anti‐indolicidin antibody were indistinguishable. © Munksgaard 1995.
AB - Indolicidin, a novel tryptophan‐rich microbicidal tridecapeptide amide isolated originally from granules of bovine neutrophils, has been prepared by optimized manual and automated protocols of stepwise solid‐phase synthesis with Nα‐9‐fluorenylmethyloxycarbonyl (Fmoc) amino acid derivatives. Both standard polystyrene (PS) and polyethylene glycol‐polystyrene (PEG‐PS) graft supports were used in combination with handles that provide C‐terminal peptide amides: 5‐(4‐Fmoc‐aminomethyl‐3,5‐dimethoxyphenoxy)valeric acid (PAL) or 5‐(9‐Fmoc‐aminoxanthen‐2‐oxy)valeric acid (XAL). Final deprotection/cleavage was carried out with reagent K, trifluoroacetic acid–phenol–water–thioanisole–1,2‐ethanedithiol (82.5:5:5:5:2.5), or reagent B, trifluoroacetic acid–phenol–water–tri(isopropyl)silane (88:5:5:2), and related cocktails. Initial purities as high as 93% were obtained immediately following cleavage. In the largest‐scale synthesis carried out, 0.8 g of HPLC‐purified indolicidin (> 99% pure) was obtained, representing a 39% overall yield based on C‐terminal Arg(Pmc) anchored to PAL‐PS‐resin. The main synthetic product, and some by‐products, were characterized by analytical high‐performance liquid chromatography (HPLC), sequencing, and fast atom bombardment mass spectrometry (FABMS). The antimicrobial potencies of natural and synthetic indolicidin, as determined by in vitro antibacterial and antifungal assays, were identical. Further, the reactivities of natural and synthetic peptides with anti‐indolicidin antibody were indistinguishable. © Munksgaard 1995.
KW - acidolytic deprotection/cleavage
KW - carbocation seavengers
KW - indolicidin
KW - solid‐phase peptide synthesis
KW - tryptophan alkylation
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U2 - 10.1111/j.1399-3011.1995.tb01055.x
DO - 10.1111/j.1399-3011.1995.tb01055.x
M3 - Article
C2 - 7591479
AN - SCOPUS:0028918846
SN - 0367-8377
VL - 45
SP - 401
EP - 409
JO - International Journal of Peptide and Protein Research
JF - International Journal of Peptide and Protein Research
IS - 5
ER -