Antifungal peptides, a heat shock protein-like peptide, and a serine-threonine kinase-like protein from Ceylon spinach seeds.
Two antifungal peptides (designated alpha- and beta-basrubrins) with molecular masses of 4-5 kDa and distinct N-terminal sequences, and a peptide and a protein with N-terminal sequences resembling heat shock protein (hsp) and serine-threonine kinase, respectively, were isolated from seeds of the Ceylon spinach Basella rubra. The purification procedure entailed saline extraction, (NH4)2SO4 precipitation, ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-cellulose, and FPLC-gel filtration on a Superdex peptide column. alpha- and beta-basrubrins inhibited mycelial growth in Botrytis cirerea with an IC50 value of 7.5 and 14.7 microM, respectively, Mycosphaerella arachidicola with an IC50 of 12.4 and 6.9 microM, and Fusarium oxysporum with an IC50 of 5.8 and 6.2 microM. Neither alpha-basrubrin nor beta-basrubin exhibited DNase, RNase, lectin or protease activity, indicating that their antifungal action is not due to these activities. HIV-1 reverse transcriptase was inhibited by alpha- and beta-basrubrins with an IC50 of 246 and 370 microM, respectively. Translation in rabbit reticulocyte lysate was inhibited by alpha- and beta-basrubrins with an IC50 of 400 and 100 nM. The heat shock protein-like peptide and serine-threonine kinase-like protein exhibited a molecular mass of 3 and 30 kDa, respectively. They inhibited neither translation in a rabbit reticulocyte system at concentrations up to 50 microM nor HIV-1 reverse transcriptase activity at concentrations up to 400 microM. They did not exert antifungal activity toward B. cinerea, M. arachidicola, and F. oxysporum when tested up to 16 microg. None of the aforementioned proteins demonstrated DNase, RNase, protease or lectin activity.
Wang
Hexiang
H
Department of Microbiology, College of Biological Science, China Agricultural University, Beijing.
Ng
Tzi Bun
TB
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000302:isolation & purification* / Q000494:pharmacology; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012639:Seeds / Q000737:chemistry*; D017421:Sequence Analysis; D018724:Spinacia oleracea / Q000737:chemistry*; D013188:Sri Lanka
D000935:Antifungal Agents; D006360:Heat-Shock Proteins; D010940:Plant Proteins; D017346:Protein Serine-Threonine Kinases