Experimental data file PPK kinetics

Selected Cell

Cell:

Value:

Kinetic measurements 31 P NMR measurements

Figure 5 D

polyP synthesis
ADP Concentration was determined discontinously with PK-LDH Assay at 37°C
Assay at 70°C	Note	PPK [µg]	ATP [mM]	Mg[mM]	TRIS Buffer [mM]	Mean specific Activity [mU/mg]	SD [mU/mg]
polyP synthesis Fig. 4D	50 mM TRIS/HCl pH 7.4 with 50 mM KCl	10	4	4	50	186.85847506068504	33.54056004743149
	PPK: heteromeric PPK	10	3	4	50	222.9983136214427	42.2830462076463
		10	2	4	50	214.4096036251298	8.691481264394412
		10	1.5	4	50	147.54072262165607	16.21818411516726
		10	1	4	50	94.36872768556537	4.2920201614784155
		10	0.75	4	50	48.33800832258233	12.79514768572167
		10	0.5	4	50	25.964760925092072	0
		10	0.25	4	50	11.212070065444362	9.272781989937625
		10	0.1	4	50	5.815863204430399	0.13905801546534644
		10	1.0E-6	4	50	0	0


ATP synthesis
ATP Concentration was determined discontinously with HK-G6PDH Assay at 37°C
Assay at 70°C
ADP in saturation	Note	PPK [µg]	polyP45 [µM]	ADP [mM]	Mg[mM]	TRIS Buffer [mM]	Mean specific Activity [U/mg]	SD [U/mg]
ATP synthesis Fig. 4E	50 mM TRIS/HCl pH 7.4 with 50 mM KCl	5	100	20	10	50	12.357054164123934	0.16987130414753718
	PPK: heteromeric PPK	5	75	20	10	50	11.912024954492345	0.2303815306587484
		5	50	20	10	50	9.137380918996133	0.13969432049383024
		5	25	20	10	50	2.825754381091484	0.08774178608431836
		5	15	20	10	50	1.2856344116727096	0.2988961645549357
		5	10	20	10	50	0.5291337894640746	0.1040952094210284
		5	7.5	20	10	50	0.2039563948923764	0.18149234330780456
		5	2.5	20	10	50	0.03708232040179955	0.16243929902202603
		5	1.0E-6	20	10	50	0	0

Assay at 70°C
polyP in saturation	Note	PPK [µg]	polyP45 [µM]	ADP [mM]	Mg[mM]	TRIS Buffer [mM]	Mean specific Activity [U/mg]	SD [U/mg]
ATP synthesis Fig. 4F	50 mM TRIS/HCl pH 7.4 with 50 mM KCl	5	100	30	10	50	2.145481576556152	0.03078524160675172
	PPK: heteromeric PPK	5	100	20	10	50	2.7641831567893242	0.019926651161207454
		5	100	10	10	50	2.7260833985444215	0.08702136113778439
		5	100	8	10	50	2.5875283391638906	0.11740873097010723
		5	100	6	10	50	2.339370500532498	0.048739013645021936
		5	100	4	10	50	2.0576400268141253	0.09302101674322075
		5	100	2	10	50	1.2699442338511673	0.10161127423823689
		5	100	1.0E-6	10	50	0	0


polyP synthesis
ADP Concentration was determined discontinously with PK-LDH Assay at 37°C
	Note	PPK [µg]	ATP [mM]	Mg[mM]	TRIS Buffer [mM]	Mean specific Activity [mU/mg]	SD [mU/mg]
ATP degradation Fig. 4F	50 mM TRIS/HCl pH 7.4 with 50 mM KCl	10	8	4	50	31.916275742245677	9.672528614336864
	PPK: homomeric PPK	10	6	4	50	35.770179787637396	10.431488660680795
		10	4	4	50	35.302105329227054	5.717076422687372
		10	3	4	50	32.59323828266209	6.134021034609912
		10	2	4	50	20.639495017595344	5.306827020552118
		10	1.5	4	50	14.123709232411553	5.155945965449727
		10	1	4	50	8.75407890829948	6.212583182219284
		10	1.0E-7	4	50	1.0E-7	0

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357

195	0.10294723623643515	0.10920886827659665	0.2976429504539443	0.1698023454261847	0.2791682257522763
210	0.10658387485457128	0.10452716300762308	0.31271600202347405	0.18069189904070398	0.2571316964890923
225	0.10491553425388118	0.0913765439144443	0.31077001747152944	0.19191276473433688	0.25588332417049753
240	0.10664581404810188	0.10539791363158028	0.303709566413621	0.20137040868880313	0.23674752371088456
255	0.1133170816103169	0.10758428424684957	0.30560315037283153	0.2087008938467002	0.22135273205676276
270	0.11854637731489963	0.107478848763884	0.3090331195665921	0.2032385653530956	0.22040856131600392
285	0.12083751969016973	0.10311871433875591	0.3060855614887555	0.22257584362962451	0.20401477334093657
300	0.13308722721504723	0.09910409301625595	0.30621454371960216	0.22864711389937684	0.18594195164739488
315	0.13594145377016326	0.09954428686536569	0.3130352492426226	0.23538797237012116	0.1664875400299924
330	0.1367356810258027	0.1090261785823659	0.3039524430701221	0.2353746010661037	0.17040027550807335
345	0.14125243476716642	0.11159566117122025	0.3093544472290164	0.24676952889581777	0.14551998706019967
360	0.14268427258581864	0.10028856613036484	0.3068033304499729	0.25171468696837096	0.14986466906777346
375	0.1415242238151172	0.10698558670502101	0.31055490289129745	0.2586076287405768	0.13535349290444434
390	0.1476572836316041	0.1203967949498569	0.29811835845360224	0.2598650884353918	0.12995737541221106
405	0.15694009088609678	0.0992921401563382	0.31150284190887584	0.28070963500993373	0.11595295371367806
420	0.15931208384686452	0.11494244253277969	0.30086877346775776	0.27397428109250915	0.10171254171749249
435	0.16143940663144335	0.10517404644477152	0.30423441317013045	0.28260536318524765	0.100558892331955
450	0.16840428069078228	0.11355908350740532	0.3030224313144858	0.2797407883169595	0.07860627069903582
465	0.1784565691125405	0.10896169693485803	0.29325459391147024	0.2979812077737165	0.07294003411039048
480	0.17575765773856877	0.11761102214129937	0.2931921670209809	0.2915042228332532	0.07148141254773137
495	0.18100822174530623	0.10784048482119457	0.30025724079928673	0.30210525490849166	0.06698318597946329
510	0.18583085598323076	0.11489243130566902	0.2881018393507894	0.2973297927406854	0.06233990312066342
525	0.19128239474481026	0.11534769354528182	0.28599252205906645	0.3165947042522441	0.040109935884650595
540	0.19150038175567338	0.11213360578808318	0.295352343871568	0.31047635174519483	0.04913773270066746
555	0.19525595823704023	0.11572589965568351	0.2859806847264849	0.30994613075465477	0.043845679412307006
570	0.20042417470810434	0.12018922835277897	0.2904901540483186	0.31708851844547054	0.031000894817127655
585	0.19289395050261482	0.1216075905099194	0.2806427714218831	0.3212450951687403	0.03770288129701838
600	0.20568261648347683	0.13061548205397855	0.27054548846602383	0.31064192149305997	0.03597063729249163
615	0.20634693836660162	0.12059455571082053	0.27038278741126504	0.33313648399200724	0.02970353907712708
630	0.20606143386624132	0.12909982206275275	0.26929939820582693	0.31564123482963335	0.03679771761797106
645	0.20961973169250434	0.12252392884990095	0.2660171358038976	0.31180546164796835	0.04271395995207238
660	0.20537951875509303	0.12526096179528304	0.26527221382752053	0.3223967549247871	0.026451434024155434
675	0.21611173135855202	0.13502833395263134	0.2618401998258388	0.3156080096760418	0.028851275760963337
690	0.22038097578995805	0.13187245990495372	0.2567719384332202	0.31774715829684397	0.030152691451066888
705	0.23126782248378697	0.1319587364633392	0.24659203079887115	0.3138455427463604	0.029534616891338128
720	0.226453008784506	0.13225865764278597	0.24826096705929118	0.31592401916813084	0.03029288403183761
735	0.22535716673864567	0.13123713366869905	0.2534279352203048	0.31471676399931536	0.034846927128097234
750	0.22993549153768217	0.13332613117987957	0.24450041997759114	0.3218025355029163	0.02558729898695544



SI Figure 12A
70°C	Note	PPK [µg]	ADP [mM]	Mg[mM]	polyP45 [µM]	TRIS Buffer [mM]	D2O [%]
ATP synthesis Fig. 5A	50 mM TRIS/HCl pH 7.4 with 50 mM KCl	0	10	10	500	50	10
	PPK: heteromeric PPK				Are responsible to get a good signal to noise ratio for polyP!
	Start by adding the enzyme
	Pi	ADP	cP3	polyP
Time [min]	Normalized sum of integrals	Normalized sum of integrals	Normalized sum of integrals	Normalized sum of integrals
0	0.0353707050752002	0.33722782436800824	0.03944675636960154	0.58795471418719
15	0.03916747294753302	0.3376490991126609	0.04494758170032114	0.5782358462394849
30	0.03481983895654792	0.34746356935605377	0.04730436909143692	0.5704122225959615
45	0.037930379238878965	0.35607514042831195	0.05324006355988371	0.5527544167729254
60	0.042403658997962584	0.3471338447312342	0.06435165981130647	0.5461108364594967
75	0.04333317813641047	0.34159164371999	0.07447088923973005	0.5406042889038695
90	0.044812570205419976	0.35537798531380943	0.0769117273229192	0.5228977171578515
105	0.04807243835167293	0.3536327117010047	0.0794221522879784	0.518872697659344
120	0.04569314492528431	0.3471762730158197	0.09670511378125801	0.5104254682776379
135	0.04504291206066771	0.353468347517076	0.09762216226142874	0.5038665781608276
150	0.04863941214575335	0.3527533236208951	0.10418119457449523	0.4944260696588563
165	0.05022099971073256	0.34508938995312016	0.11479623886920251	0.4898933714669448
180	0.05272509275138556	0.3465398570648486	0.12130848235783889	0.479426567825927
195	0.052997373771801075	0.35105282031730023	0.1281119000437742	0.4678379058671245
210	0.05589054780021302	0.3548219464655944	0.13326945949062835	0.45601804624356423
225	0.06343085301030033	0.3467870772381352	0.14128277025709704	0.44849929949446743
240	0.05836904229463555	0.36394731239435957	0.14058622116309646	0.4370974241479084
255	0.06874154856055625	0.35328130864604745	0.1486376052646481	0.4293395375287483
270	0.0681870169233257	0.35007978689943803	0.15892112170094183	0.4228120744762946
285	0.06627338870317974	0.3653651124193105	0.16358276966358834	0.4047787292139214
300	0.07074184993425933	0.3470675848802718	0.17154619957444475	0.4106443656110242
315	0.0710740276331569	0.35159906204873226	0.17888677107843473	0.39844013923967614
330	0.07149053237423666	0.3614979591004233	0.1802605742354069	0.3867509342899332
345	0.07990608647471772	0.34992300763904594	0.19159280580164725	0.37857810008458925
360	0.07598589549654587	0.3565799591453446	0.19774964571481243	0.36968449964329697
375	0.08091002648628878	0.35303589831874954	0.2031508172552938	0.36290325793966793
390	0.08329364563049627	0.35339412793379504	0.20964861774606924	0.3536636086896395
405	0.08216172974440415	0.3580908634668763	0.21524878793039565	0.34449861885832384
420	0.08734214449423726	0.35613627535558134	0.22439444313181914	0.33212713701836216
435	0.08171463883430011	0.35158210653554167	0.23183880249832967	0.3348644521318285
450	0.09130821073731551	0.3573090276039173	0.23101445557878872	0.32036830607997846
465	0.09296780452415318	0.3575288991618176	0.24033068411946462	0.30917261219456466
480	0.09225964671706095	0.35881869150331536	0.2429352298986293	0.3059864318809944
495	0.09540080646227968	0.35829664202928546	0.24335570306673115	0.30294684844170366
510	0.10441914029460589	0.34907803363751805	0.2539961996533093	0.29250662641456676
525	0.09571764654567513	0.3489473653309947	0.2667018092622	0.28863317886113016
540	0.10142610189364334	0.3560238461188505	0.26961363690033785	0.27293641508716837
555	0.10436404452046419	0.36132733812467815	0.2747412542778053	0.2595673630770523
570	0.10472331200328848	0.34904266601264855	0.283438458173366	0.2627955638106971
585	0.11189589882592284	0.35206740994013397	0.2855361968699634	0.25050049436397986
600	0.11195325338328159	0.3448051073876553	0.2921109685444674	0.2511306706845958
615	0.1136492510496457	0.3490877037981478	0.2961874429245334	0.241075602227673
630	0.11310203158914789	0.3559688519631312	0.29939696545467276	0.23153215099304816
645	0.11736433221903174	0.3509871876158716	0.3068800601820235	0.22476841998307318
660	0.12095108369794416	0.34975000044709903	0.3058975098336856	0.22340140602127112
675	0.11800474805189126	0.353455641122999	0.31089431006836277	0.2176453007567471
690	0.11697114794558706	0.3475411374006382	0.3199791817992833	0.2155085328544916
705	0.12081365175020417	0.3527497610329829	0.3278809515125283	0.19855563570428472
720	0.11989564141263713	0.34443461040136597	0.3271931995290024	0.20847654865699453


SI Figure 12C
70°C	Note	homomeric PPK [µg] (Saci_2019)	ATP [mM]	Mg[mM]	PEP [mM]	Sso_PK [µg]	TRIS Buffer [mM]	D2O [%]
ATP synthesis Fig. 5A	50 mM TRIS/HCl pH 7.4 with 50 mM KCl	20	2	20	10	8	50	10
	PPK: heteromeric PPK
	Start by adding the enzyme
	Pi	PEP	ATP		No formation of polyp detectable
Time [min]	Normalized sum of integrals	Normalized sum of integrals	Normalized sum of integrals
0	0	0.8411189934413064	0.1588810065586936
15	0.033342309612848106	0.7981351612204627	0.16852252916668917
30	0.03677706845603591	0.7979134512249411	0.16530948031902296
45	0.04453025883130216	0.805681911630274	0.14978782953842382
60	0.04828813069924583	0.8059789504115858	0.14573291888916834
75	0.06070095449946654	0.7810985577802866	0.15820048772024686
90	0.07997489991348816	0.7555764684269972	0.16444863165951454
105	0.09089035586690751	0.7558377213968241	0.1532719227362682
120	0.09735258983799924	0.7456215300262176	0.1570258801357832
135	0.09060448918229548	0.7592766468938607	0.1501188639238438
150	0.09609209961607951	0.7358164034511213	0.1680914969327992
165	0.11926358050266432	0.7127841270946033	0.16795229240273243
180	0.13366631796332937	0.7014478948531225	0.164885787183548
195	0.1372116259899652	0.6966130265955142	0.16617534741452059
210	0.13927151545286734	0.6923077471030683	0.1684207374440645
225	0.14965056615014327	0.6882465468117217	0.162102887038135
240	0.1448867148500572	0.690481846691744	0.1646314384581989
255	0.16564120023692339	0.676610814141562	0.15774798562151465
270	0.16395348670290805	0.6727910696330353	0.16325544366405653
285	0.18401217113226867	0.6525062259922388	0.16348160287549232
300	0.17490377478789843	0.6474096061884126	0.17768661902368887
315	0.19656040911431497	0.6252572756453686	0.17818231524031652
330	0.20223618653451064	0.6232864037262449	0.17447740973924442
345	0.22077468643515832	0.6201415821753206	0.15908373138952112
360	0.21668039929600652	0.6060754244102907	0.17724417629370273
375	0.23204236489808588	0.604325544641918	0.16363209045999597
390	0.23342042002035782	0.6020618122091496	0.16451776777049257
405	0.24224432369866902	0.5873082374672729	0.17044743883405813
420	0.2407663584461979	0.5780113969500653	0.18122224460373676
435	0.2519505769463104	0.5713467010149416	0.176702722038748
450	0.25267396823922555	0.575708852214419	0.17161717954635533
465	0.2642517753044398	0.5602489239018036	0.17549930079375667
480	0.25693658572719585	0.5577822327542485	0.1852811815185557
495	0.2686860642292029	0.5550823193552934	0.17623161641550386
510	0.2919885131403755	0.5461111989140905	0.161900287945534
525	0.2874734668508136	0.5323643935826636	0.1801621395665226
540	0.290578250435139	0.5509502196390104	0.1584715299258506
555	0.304802047631471	0.5159650233083352	0.17923292906019384
570	0.3086928518799503	0.5143199028590905	0.1769872452609592
585	0.3025268219904718	0.5122477438074005	0.1852254342021277
600	0.325422588374074	0.505502050822209	0.16907536080371693
615	0.3423179011962227	0.49033511198295615	0.16734698682082125
630	0.3361242510282669	0.48698150120320255	0.17689424776853058
645	0.33649666795345007	0.4904840987055104	0.1730192333410395
660	0.3412226473270425	0.46507741039677486	0.19369994227618267
675	0.3636312026409448	0.46575251882942237	0.17061627852963296
690	0.34521987083254735	0.47016597658445325	0.18461415258299946
705	0.3559456463717811	0.4746194565585924	0.16943489706962647
720	0.3701521561297833	0.46087845496056795	0.1689693889096487
735	0.37317719440377933	0.43770686485930815	0.1891159407369125
750	0.3635944685550884	0.45542132176266015	0.18098420968225132