Purification and characterization of a novel salivary antimicrobial peptide from the tick, Ixodes scapularis.
A novel antimicrobial peptide was isolated from the saliva of the Lyme disease tick vector, Ixodes scapularis, henceforth designated as ISAMP (I. scapularis Antimicrobial Peptide). ISAMP was purified using a sequential method including ultra filtration, gel filtration and reverse-phase high performance liquid chromatography. The purified peak had a molecular weight of 5.3kDa by MALDI/TOF-MS and its amino acid sequence, determined by Edman degradation was PDxGxPxxVKAGRxPxxSI. A BLASTP search revealed that the protein is a putative 5.3kDa secreted protein (AAM93656) from I. scapularis. The predicted protein is composed of 69 amino acids with no conserved domain motifs. Purified ISAMP was found to have antimicrobial activities against bacteria. Gene expression studies were carried out to observe ISAMP expression in different tick tissues. RT-PCR results indicated that the gene was expressed in hemocytes, fat body and salivary gland but virtually no expression was observed in the midgut. ISAMP is only similar to other Ixodid tick proteins, thus it is a member of a unique family.
Center for Vector-Borne Disease, University of Rhode Island, Kingston, RI 02881, USA.
Sivakamasundari
S
Pichu
José M C
JM
Ribeiro
Thomas N
TN
Mather
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000187:drug effects; D053878:Hemolytic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D018884:Ixodes / Q000378:metabolism*; D012471:Salivary Proteins and Peptides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D014462:Ultrafiltration
D023181:Antimicrobial Cationic Peptides; D053878:Hemolytic Agents; D012471:Salivary Proteins and Peptides