Purification and antimicrobial activity studies of the N-terminal fragment of ubiquitin from human amniotic fluid.
A 4.3-kDa antimicrobial peptide was isolated from human amniotic fluid by dialysis, ultrafiltration, and C18 reversed-phase high performance liquid chromatography. This peptide, which we named Amniotic Fluid Peptide-1 (AFP-1), possessed antimicrobial activity but lacked hemolytic activity. In addition, AFP-1 potently inhibited the growth of a variety of bacteria (Escherichia coli, Salmonella typhimurium, Listeria monocytogenes and Staphylococcus aureus), filamentous fungi (Botrytis cinerea, Aspergillus fumigatus, Neurospora crassa and Fusarium oxysporum) and yeast cells (Candida albicans and Cryptococcus neoformans). Automated Edman degradation showed that the N-terminal sequence of AFP-1 was NH(2)-Met-Gln-Ile-Phe-Val-Lys-Thr-Leu-Thr-Gly-Lys-Thr-Ile-Thr-Leu-Glu-Val-Glu-. The partial sequence had 100% homology to the N-terminal sequence of ubiquitin. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that the molecular mass of AFP-1 was 4280.2 Da. Our data show an antimicrobial activity of ubiquitin N-terminal derived peptide that makes it suitable for use as an antimicrobial agent.
Kim
Jin-Young
JY
Research Center for Proteineous Materials, Chosun University, Kwangju 501-759, Korea.
Lee
Sun Young
SY
Park
Seong-Cheol
SC
Shin
Song Yub
SY
Choi
Sang Joon
SJ
Park
Yoonkyung
Y
Hahm
Kyung-Soo
KS
D000595:Amino Acid Sequence; D000653:Amniotic Fluid / Q000737:chemistry*; D000890:Anti-Infective Agents / Q000302:isolation & purification* / Q000494:pharmacology*; D005260:Female; D006801:Humans; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification* / Q000494:pharmacology*; D011247:Pregnancy; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D025801:Ubiquitin / Q000302:isolation & purification* / Q000494:pharmacology
D000890:Anti-Infective Agents; D010446:Peptide Fragments; D025801:Ubiquitin; C523872:amniotic fluid peptide-1, human