During the assembly of the bacteriophage T4 prehead, a T4-coded protease zymogen (P21) is built into the structure. At a certain stage in head formation, the protease precursor is activated and specifically cleaves most of the prehead proteins. In this paper we show that a correlation existed between the presence of proteinaceous material in the center of the prehead core, observed by electron microscopy, and the availability of P21 during prehead assembly. In the absence of P21, the core enclosed a hold of about 35 nm long and 20 nm wide. We found the same for (i) in vitro-assembled, negatively stained prehead-like structures and (ii) in vivo-formed preheads in thin sections of T4-infected cells. We concluded that P21 was localized in the center of the prehead core.
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