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Figure 1.
(a) N-terminal sequences of R^21, Lyz^P1 and T4 E. The
N-terminal domains of R^21, Lyz^P1 and T4 E are shown aligned by
their Glu-8aa-Asp/Cys-5aa-Thr catalytic triad (blue and
asterisks). SAR domains are boxed in orange. Leucine
substitutions made in R^21 are shown above Gly14 and Gly15. The
PelB signal sequence (PelB[ss]) and the artificial transmembrane
domain (TMD[art]; yellow) are shown with arrows indicating
points of fusion in the chimeric constructs. Residues involved
in positioning the catalytic glutamate are highlighted by a
light blue box in Lyz^P1 and T4 E. (b) Lysis profiles. (  )
pZE-luc, (  circle
) pZE-R^21[G14,15L], (  )
pZE-luc + 1 mM CHCl[3] at 80 min after induction, (  square
) pZE-R^21[G14,15L] + 1 mM CHCl[3] at 80 min after induction, (
 )
pZE-R^21, (  diamond
)Lyz^P1[1–26]  R^21[27–165],
(  )
pZE-pelB[ss] R^21[27–165],
(  triangle
) pZE-TMD[art] R^21.
A[550], absorbance at 550 nm. (c) Localization and processing of
R^21 and derivatives after expression of the indicated chimeras.
In top panel, lanes 1, 2 and 3 represent the total (T),
periplasm (P) and spheroplast (S) fractions, respectively.
Below, lanes 1, 2 and 3 represent the total (T), soluble (S) and
membrane (M) fractions, respectively. (d) Morphologies of cells
expressing the indicated pZE-R^21 plasmids at 100 min after
induction. Scale bars (throughout) are 5  m.
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