Purification and characterization of the lysozyme from the gut of the soft tick Ornithodoros moubata.
The gut of the adult soft ticks Ornithodoros moubata displays high lytic activity against the bacteria Micrococcus luteus. The activity differed in the range of two orders of magnitude among individual animals and increased on average 4 fold during the first week following ingestion. In homogenates of first instar nymphs the activity was much lower increasing exponentially as nymphs neared the first molt. The protein responsible for this activity was purified out of gut contents of adult ticks by means of affinity adsorption on magnetic-chitin followed by two chromatography steps on cation exchange FPLC column MonoS. The homogeneous active protein has a mass of 14006 +/- 20 Daltons as determined by MALDI-TOF mass spectrometry. The N-terminal amino-acid sequence of this protein is K-V-Y-D-R-C-S-L-A-S-E-L-R with the highest similarity to the lysozyme from liver of rainbow trout and to lysozymes from digestive tracts of several mammals. The motif DRCSLA is specific for the digestive lysozymes of several dipteran insects. Based on this evidence, we have identified the protein as the tick gut lysozyme. The tick gut lysozyme has a pI near 9.7 and retains its full activity after treatment at 60 degrees C for 30 minutes. The pH optimum of the tick lysozyme was in the range from pH 5-7. Only marginal activity could be detected at pH > 8 which raises the question about the function of lysozyme in anti-bacterial defense in the environment of the tick gut.
Kopácek
P
P
Institute of Parasitology of the Czech Academy of Sciences, Ceské Budĕjovice, Czech Republic. kopajz@paru.cas.cz
Vogt
R
R
Jindrák
L
L
Weise
C
C
Safarík
I
I
D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000187:drug effects; D002845:Chromatography; D004064:Digestive System / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009113:Muramidase / Q000302:isolation & purification* / Q000378:metabolism; D013987:Ticks / Q000201:enzymology*
D009113:Muramidase