1887

Abstract

The Gram-positive eubacterium contains five chromosomally encoded type I signal peptidases (SPases) for the processing of secretory pre-proteins. Even though four of these SPases, denoted SipS, SipT, SipU and SipV, are homologous to the unique SPase I of , they are structurally different from that enzyme, being almost half the size and containing one membrane anchor instead of two. To investigate whether the unique membrane anchor of SPases is required for activity, soluble forms of SipS of , SipS of and SipC of the thermophile were constructed. Of these three proteins, only a hexa-histidine-tagged soluble form of SipS of could be isolated in significant quantities. This protein displayed optimal activity at pH 10, which is remarkable considering the fact that the catalytic domain of SPases is located in an acidic environment at the outer surface of the membrane of living cells. Strikingly, in contrast to what has been previously reported for the soluble form of the SPase, soluble SipS was active in the absence of added detergents. This observation can be explained by the fact that a highly hydrophobic surface domain of the SPase, implicated in detergent-binding, is absent from SipS.

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2001-04-01
2020-01-26
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