Structure, stability, and biological activity of bacteriophage T4 gene product 9 probed with mutagenesis and monoclonal antibodies

Journal of Structural Biology
L P KurochkinaVadim V Mesyanzhinov

Abstract

Gene product (gp) 9 connects the long tail fibers and triggers the structural transition of T4 phage baseplate at the beginning of infection process. Gp9 is a parallel homotrimer with 288 amino acid residues per chain that forms three domains. To investigate the role of the gp9 amino terminus, we have engineered a set of mutants with deletions and random substitutions in this part. The structure of the mutants was probed using monoclonal antibodies that bind to either N-terminal, middle, or C-terminal domains. Deletions of up to 12 N-terminal residues as well as random substitutions of the second, third and fourth residues yielded trimers that failed to incorporate in vitro into the T4 9(-)-particles and were not able to convert them into infectious virions. As detected using monoclonal antibodies, these mutants undergo structural changes in both N-terminal and middle domains. Furthermore, deletion of the first twenty residues caused profound structural changes in all three gp9 domains. In addition, N-terminally truncated proteins and randomized mutants formed SDS-resistant "conformers" due to unwinding of the N-terminal region. Co-expression of the full-length gp9 and the mutant lacking first 20 residues clearly shows the asse...Continue Reading

References

Sep 1, 1992·Bioscience, Biotechnology, and Biochemistry·A KatoI Kumagai
Aug 1, 1991·The Journal of Cell Biology·I BraakmanA Helenius
Oct 1, 1991·Trends in Biochemical Sciences·M E Goldberg
Jan 1, 1990·Methods in Enzymology·F W StudierJ W Dubendorff
Mar 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·S Blond, M Goldberg
Jan 1, 1989·Annual Review of Biochemistry·M G Rossmann, J E Johnson
Mar 1, 1966·Proceedings of the National Academy of Sciences of the United States of America·R S Edgar, W B Wood
Feb 25, 1980·Journal of Molecular Biology·R A Crowther
Dec 1, 1995·Protein Science : a Publication of the Protein Society·M J BennettD Eisenberg
Jan 1, 1997·Protein Science : a Publication of the Protein Society·M A SpeedJ King
Jan 1, 1997·Advances in Protein Chemistry·M P SchluneggerD Eisenberg
Feb 27, 1999·Current Opinion in Structural Biology·L Liljas
Mar 23, 2002·Protein Science : a Publication of the Protein Society·Jason F KreisbergJonathan King
Aug 19, 2003·Nature Structural Biology·Victor A KostyuchenkoMichael G Rossmann
Nov 25, 2003·Protein Science : a Publication of the Protein Society·Matthew J Gage, Anne Skaja Robinson
Aug 24, 2005·Nature Structural & Molecular Biology·Victor A KostyuchenkoMichael G Rossmann

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Citations

Nov 4, 2008·Biochemistry. Biokhimii︠a︡·L P KurochkinaV V Mesyanzhinov
Jan 4, 2021·Current Opinion in Biotechnology·Katarzyna Gembara, Krystyna Dąbrowska

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