Effect of the viral protease on the dynamics of bacteriophage HK97 maturation intermediates characterized by variance analysis of cryo EM particle ensembles

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• 作者：Yunye Gong^a ; ^{yg326@cornell.edu" class="auth_mail" title="E-mail the corresponding author} ; David Veesler^b ; ¹ ; ^{dveesler@uw.edu" class="auth_mail" title="E-mail the corresponding author} ; Peter C. Doerschuk^c ; ^{pd83@cornell.edu" class="auth_mail" title="E-mail the corresponding author} ; John E. Johnson^b ; ^{jackj@scripps.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Cryo electron microscopy ; Virus maturation ; Hong Kong 97 bacteriophage ; Viral protease
• 刊名：Journal of Structural Biology
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：193
• 期：3
• 页码：188-195
• 全文大小：2165 K

文摘

Cryo EM structures of maturation-intermediate Prohead I of bacteriophage HK97 with (47847715301180&_mathId=si1.gif&_user=111111111&_pii=S1047847715301180&_rdoc=1&_issn=10478477&md5=e0e45644844fa668f09cb7b92b5cd649" title="Click to view the MathML source">PhI^Pro+${\text{PhI}}^{\text{Pro+}}$) and without (47847715301180&_mathId=si2.gif&_user=111111111&_pii=S1047847715301180&_rdoc=1&_issn=10478477&md5=97218720c76c4a26e3d55217bba597af" title="Click to view the MathML source">PhI^Pro−${\text{PhI}}^{\text{Pro−}}$) the viral protease packaged have been reported (Veesler et al., 2014). In spite of 47847715301180&_mathId=si1.gif&_user=111111111&_pii=S1047847715301180&_rdoc=1&_issn=10478477&md5=e0e45644844fa668f09cb7b92b5cd649" title="Click to view the MathML source">PhI^Pro+${\text{PhI}}^{\text{Pro+}}$ containing an additional 47847715301180&_mathId=si36.gif&_user=111111111&_pii=S1047847715301180&_rdoc=1&_issn=10478477&md5=7c231ff225aa8407a501845b93f53c63">47847715301180-si36.gif">$\sim 100\times 24\text{kD}$ of protein, the two structures appeared identical although the two particles have substantially different biochemical properties, e.g., 47847715301180&_mathId=si2.gif&_user=111111111&_pii=S1047847715301180&_rdoc=1&_issn=10478477&md5=97218720c76c4a26e3d55217bba597af" title="Click to view the MathML source">PhI^Pro−${\text{PhI}}^{\text{Pro−}}$ is less stable to disassembly conditions such as urea. Here the same cryo EM images are used to characterize the spatial heterogeneity of the particles at 17 Å resolution by variance analysis and show that 47847715301180&_mathId=si2.gif&_user=111111111&_pii=S1047847715301180&_rdoc=1&_issn=10478477&md5=97218720c76c4a26e3d55217bba597af" title="Click to view the MathML source">PhI^Pro−${\text{PhI}}^{\text{Pro−}}$ has roughly twice the standard deviation of 47847715301180&_mathId=si1.gif&_user=111111111&_pii=S1047847715301180&_rdoc=1&_issn=10478477&md5=e0e45644844fa668f09cb7b92b5cd649" title="Click to view the MathML source">PhI^Pro+${\text{PhI}}^{\text{Pro+}}$. Furthermore, the greatest differences in standard deviation are present in the region where the δ-domain, not seen in X-ray crystallographic structures or fully seen in cryo EM, is expected to be located. Thus presence of the protease appears to stabilize the δ-domain which the protease will eventually digest.