Original title: Studium regulace NADP - malátdehydrogenasy (dekarboxylační) a vliv stresu na tento enzym
Translated title: Study of the Regulation of NADP-Malic Enzyme (Decarboxylating) and Influence of Stress on this Enzyme
Authors: Doubnerová, Veronika
Document type: Rigorous theses
Year: 2009
Language: cze
Abstract: [cze] [eng]
4. Závér l. Z listůtabiůal Nicotiana tabacum L. byla izolovrfura nefotosyntetická cýosolová isoforma NADP-dependentní matátdehydÍogenasy (oxaloacetát-dekarboxy|ační) (EC l.l.l.a.; NADP-ME) o výsledné specifické aktivitě 0,95 pm'ol.miďI.mgl, rolďivní molekulové hmotnosti jedné podjednotky tetameru - 67 0oo, pI 5,5 a pH optimu 7,| - 7,4. By|y stanoveny kinetické parametry tétoisoformy pro subshát L-malfu a NADP+ v pfftomnosti kofaktorů tutď-' u"1 co1 Ni1 stejně jako byly sledovríny záúslosti reakčnich rychlostí na koncentaci jednotlivých iontů. Závislost reakčnírychlosti na koncentraci Mn2* iontů byla signoi&írníse silnou pozitivní kooperďivitou a vysokou hodnotou Hillova koefi cientu 7,5. -10- 2. Mechanismus dvousubstnítové reakce katalyzované Njako uspořádaný sekverrční. IADP-ME z listů tabríku by| určen 3. Regulace NADP-ME isoÍbrmy purifikované z listů energeticky boha!Ími lát g|yko|ýZy. kami (cTP, ATP a ADP)' 4. Vlivem biotického s[ ke anýšení.kti"ity ;:;í;.TTH::,í::r"*@VY) došto vprůběhu infekce kmenem PVY1 tat *;;""',:Jfr".:,:":::l i kořenech N. fubacum L.' a to jak aktivity než ".*",,.u'1j";J;:il#'fiT kmen PVY. způsobil nixi ',"ys"ni #:,:;il";"1y'-;;#;;'"f, il.:'3T]"",;;1ňTť: 5. Transgenní rost|iny Nicotiana bbacuml. s vnesen.. z A viru bramboru a tt m u| ti fu nkčnío",,"ř" Jř:':... 4. Conclusions l. Non-photosynthetic cytosolic isoform of NADP-mďic ilZ-oe (oxaloacetate decarboxylating) (NADp-ME; EC r.l.l.4o) was isolated from tobacco (Nicoriana tabacun L.) leaves. Specific aďiviý of the obtained enz5me preparation was 0.95pmol.miď|.mg l, relďive molecular mass of one subr'nit of tetramer approximatety 67000' pI 5'5 and pH optimum 7.r - 7.4. Kinetic constants of this isoform were determined for L-ma|ate and NADP+ in the presence of various cofactors Mď*, Mo*,co2', Ni2*, similarly the dependencies ofreaction Íates on the ooncentration ofdivalent metal ions were observed- The dependence of reaction rate on Mn2+ concentration wassigmoidal with strong positive cooperativiý and high value of Hill coefficient 7.5. -2r - J. 4. Summarv ofPh.D.Thesis 2. The kinetic meohanism of two-substate reaďion catalyzed by NADP.ME ůomtobacco leaves was determined as ordered sequmtial. The regulation of the NADP-ME purified ůomtobacco leaves occurs predominant|y in the prsence of macmergic compounds (GTP, ATP and ADP), sligbtly by metabolites of glymlysis. Biotic stress caused by potato yirus f G1;Y) (not only strain PWNN but also PWo) increased NADP.ME activiý in N. tabacum L. |eaves, stems and roots during the infeďion. Milder PVYo caused tower increase of NADP-ME aďivity than necrotic isolate of PVYNN....

Institution: Charles University Faculties (theses) (web)
Document availability information: Available in the Charles University Digital Repository.
Original record: http://hdl.handle.net/20.500.11956/93739

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