BCHE 395-12 Enzyme Kinetics I

BCHE 395-12 Enzyme Kinetics I - Enzyme Kine*cs Study of...

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Enzyme’Kine*cs’ Study’of’factors’that’aFect’the’ rates ’of’ enzyme-catalyzed’reac*ons’ Knowing’the’kine*c’behavior’of’enzymes’ under’various’condi*ons’allows’one’to’fully’ characterize’the’nature’of’the’enzyme’ O?en’kine*cs’is’used’to’determine’the’ mechanism of catalysis
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Enzyme’Kine*cs’ Kine*cs’can’provide’evidence’for:’ Reac*on’intermediates’ The’order’in’which’reactants’and’products’ leave’the’ac*ve’site’ Intracellular’concentra*ons’of’reactants’ Regulatory’mechanisms’ The’iden*ty’of’the’amino’acid’R-groups’of’the’ ac*ve’site’
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Important’Defni*ons ’ ’ Rate (velocity à V ): the’amount’oF’substrate’that’ reacts’per’unit’*me’( e.g:’s -1 ,’min -1 )’ Rate law: describes’how’the’rate’is’depends’on’the’ concentra*on’oF’a’substrate ’ ’ V ’=’ k [S] V = k [S 1 ][S 2 ]’ Rate order :’sum’oF’the’power’oF’concentra*on’in’the’ rate’law’expression’with’respect’to’the’reactants’ The’number’oF’di±erent’molecules’than’need’to’collide’ simultaneously’to’react’ V max = maximal rate of the reac±on 1 st ’order’ 2 nd ’order’
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Enzyme’Kine*cs:’ Simple,’First’Order’Chemical’Reac*ons Start’with’rates’of’ frst order chemical reac*on’ S’ à ’P’with’reac*on’rate’constant’ k k ’=’1/*me Reac*on’rate’is’dependent’ONLY’on’the’ concentra*on’of’ one ’reactant’[S]’ V ’=’ k [S]’
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Enzyme’Kine*cs:’S’ ’P ’ Since’ V ’is’ dependent’on’[S]’ we’can’measure’ rates’at’diferent’ [S]’ Problem:’[S]’is’ changing’through’ the’assay’ à ’gets’ used’up! ’ So, need to plot ini:al rates vs. [S] V 0 = ini:al velocity Lines’are’the’ V ’(velocity’oF’reac*on)’
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Enzyme’Kine*cs:’S’ ’P’ V 0 vs.’[S]’ [E]’constant’ Vary’[S]’ à Measure’ V 0 ’and’ plot’ V max ’is’plateau’ So’much’[S],’ enzyme’working’at’ maximum’velocity’ w w w w w w w
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BCHE 395-12 Enzyme Kinetics I - Enzyme Kine*cs Study of...

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