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Struktura in dinamika glavnega plaščnega proteina bakteriofaga M13 : magistrsko delo = Structure and dynamics of bacteriophage M13 major coat protein : master of science thesisStopar, DavidGlavni plaščni protein bakteriofaga M13 je del reprodukcijskega cikla vstavljen v membrano gostitelja. Strukturno je sestavljen iz dolgega hidrofobnega heliksa in krajšega N-končnega heliksa na ... površini membrane, ki je pravokoten na normalo membrane. Oba konca proteina, C- in N- sta zelo mobilna in nista inkorporirana v membrano. Pri nastanku novega virusnega delca v membrani gostitelja, prihaja do znatnih strukturnih sprememb glavnega plaščnega proteina. Da bi opisali dinamiko N-terminalnega konca proteina smo spinsko označili terminalno amino skupino glavnega plaščnega proteina v intaktnem bakteriofagu. Zaradi nespecifičnega spinskega označevanja intaktnega virusnega delca smo v primarni strukturi proteina mesto-specifično vstavili enojne cisteinske aminokislinske ostanke. Glavni kriterij za vse mutante je bila njihova sposobnost zaključevanja reproduktivnega cikla. Tri mutante v C-končnem delu proteina (G38C, T46C in A49C) smo nadalje derivatizirali z maleimidnim spinskim označevalcem in rekonstituirali v kolatne, tritonske in CTAB micele. Dinamika z lizini bogatega C-terminalnega konca proteina rekonstituiranega v micele kaže, da lahko naboji glavnega plaščnega proteina igrajo pomembno vlogo pri vzdrževanju stabilne termodinamske asociacije med proteini in membrano. Čeprav so bili v membrano rekonstituirani mutirani plaščni proteini strukturno enaki kot nemutirani proteini se je pridelek mutiranih bakteriofagov signifikantno zmanjšal. To skupaj z majhno viabilnostjo mutantov (sedem viabilnih mutantov od skupno devetnajstih), kaže da je primarna sekvenca glavnega plašča proteina zelo konzervativna in dovoljuje le manjše spremembe. Nakazana je tudi možnost nadaljnih raziskav.Type of material - master's thesis ; adult, seriousPublication and manufacture - Ljubljana : [D. Stopar], 1995Language - slovenianCOBISS.SI-ID - 12151
Author
Stopar, David
Other authors
Megušar, France
Topics
bakteriofagi |
M13 |
Escherichia coli |
plaščni proteini |
fosfolipidne membrane |
krožni dihroizem |
ESR - elektronska spinska resonanca |
mesto specifični mutanti |
spinsko označevanje |
spinsko označevanje
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Database name | Field | Year |
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Stopar, David | 13005 |
Megušar, France | 00529 |
Source: Personal bibliographies
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