E. coli o-spanin, also known as Outer membrane lipoprotein Rz1 from lambdoid prophage DLP12, RzoD or RzoR, is originated from the defective Coliphage lambda DLP12 genome carried by E. coli and present in nearly all lambdoid phages and many additional bacteriophages. DLP12 cassette in E. coli contains four lysis proteins ORFs; a holin (S) through which the endolysin (R) and spanin (Rz/Rz1) escape into the periplasmic space. Two Spanin cell-lysis proteins, o-spanin an outer membrane lipoprotein and i-spanin (rzpD) an integral inner membrane protein are homologues of the Lambda phage RZ1 and RZ genes respectively, the Rz1 gene is uniquely located within the +1 reading frame of Rz.
Specificity
Anti-o-spanin antibody specifically recognizes the Outer membrane lipoprotein subunit o-spanin from E. coli and Shigella flexneri and does not cross react with Proteus mirabilis.
Immunogen
Peptide: E.coli O-spanin. CAWIMQPPPDWQTPLNG
Application
The antibody may be used in various immunochemical techniques including Immunoblotting (predicted ~6.5kDa observed ~20 kDa due to oligomerization). Detection of the o-spanin band by Immunoblotting is specifically inhibited by the immunizing peptide.
Biochem/physiol Actions
Spanins are conserved among lambdoid phages and comprise the final step of lysis in lambda phage infections of E. coli. These proteins form heterotetramer complex composed of two homodimers linked by homotypic intermolecular disulfide bonds, span the periplasm and bridge the cell envelope thus causing fusion of the inner and outer membranes and disruption of the bacterium membrane.The DLP12 lysis genes play an important, yet to be identified role in the bacterial cell wall maintenance and biofilm formation. This was shown using mutated strains lacking essD, ybcS and rzpD/rzoD that were unable to form biofilms and had significant lower aggregation as compared with wild-type. These finding suggests that defective prophage genes are affecting host cell physiology contribute to host fitness and survival, biofilm dispersion, enhanced biofilm traits such as antibiotic resistance, attachment, phage tolerance and metabolite recycling.
Physical form
Solution in 0.01 M phosphate buffered saline pH 7.4, containing 15 mM sodium azide as a preservative.
Storage and Stability
For continuous use, store at 2-8°C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.
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