Post on 18-Dec-2015
StudyStudy of Complex of Complex Associates Between Associates Between
Cationic Porphyrins and Cationic Porphyrins and Nucleosides or Nucleosides or
NucleotidesNucleotides
Magdalena MakarskaMagdalena Makarska, Stanisław Radzki, Stanisław Radzki
Department of Inorganic ChemistryDepartment of Inorganic Chemistry
Maria Curie-Skłodowska University in LublinMaria Curie-Skłodowska University in Lublin
HH22TMePyPTMePyP5,10,15,20-tetrakis[4-(1-methyl-4-5,10,15,20-tetrakis[4-(1-methyl-4-
pyridyl)]porphyrinpyridyl)]porphyrin
N
N
N
N
N N
NN
CH3
CH3
CH3
CH3
+ +
+
+ H3C
4
H
H
SO3
-
CC4444HH3838NN88 · 4(C · 4(C77HH77SOSO33) = 1363.63 g/mol) = 1363.63 g/mol
violet powder, dark purple in solutionviolet powder, dark purple in solution
HH22TTMePPTTMePP5,10,15,20-tetrakis[4-5,10,15,20-tetrakis[4-
(trimethylamino)phenyl]porphyrin(trimethylamino)phenyl]porphyrin
N N
NN
CH3
+
+
H
H
-H3C
4
SO3
NCH3 CH3
CH3
+
NCH3
CH3
CH3
+
N
CH3
CH3
CH3
N
CH3
CH3
CC8484HH9090NN88OO1212SS44 = 1531.95 g/mol = 1531.95 g/mol
dark red powder, red in solutiondark red powder, red in solution
H2TMePyP, [H2O], cuvette 1 cmH2TTMePP, [H2O], cuvette 1 cm
[nm]200 250 300 350 400 450 500 550 600 650 700 750 800 850 900
A
0.00
0.25
0.50
0.75
1.00
1.25
1.50
1.75
2.00
500 550 600 650 700 750
0.00
0.02
0.04
0.06
0.08
0.10
0.12
423
414
520, 556, 586, 643
516, 551, 579, 636
Molar absorbancy index for H2TMePyP423 nm - 1.5680520 nm - 0.1045556 nm - 0.0411586 nm - 0.0463643 nm - 0.0135
Molar absorbancy index for H2TTMePP
414 nm - 1.8646516 nm - 0.0706551 nm - 0.0253
579 nm - 0.02752636 nm - 0.0139
Q bandSoret band
H2TMePyP, [H2O], cuvette 1 cmH2TTMePP, [H2O], cuvette 1 cm
[nm]200 250 300 350 400 450 500 550 600 650 700 750 800 850 900
A
0.00
0.25
0.50
0.75
1.00
1.25
1.50
1.75
2.00
500 550 600 650 700 750
0.00
0.02
0.04
0.06
0.08
0.10
0.12
423
414
520, 556, 586, 643
516, 551, 579, 636
Molar absorbancy index for H2TMePyP423 nm - 1.5680520 nm - 0.1045556 nm - 0.0411586 nm - 0.0463643 nm - 0.0135
Molar absorbancy index for H2TTMePP
414 nm - 1.8646516 nm - 0.0706551 nm - 0.0253
579 nm - 0.02752636 nm - 0.0139
Q bandSoret band
Molar absorbancy Molar absorbancy indexesindexes
Cu(ac)2 + H2TMePyP
Cu(ac)2 + H2TTMePP
[nm]
200 250 300 350 400 450 500 550 600 650 700 750 800 850 900
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4A
550
426
413
540
Cu(ac)2 + H2TMePyP
Cu(ac)2 + H2TTMePP
[nm]
200 250 300 350 400 450 500 550 600 650 700 750 800 850 900
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4A
550
426
413
540
Spectra of porphyrin complexesSpectra of porphyrin complexes with copper (II)with copper (II)
Protonation of HProtonation of H22TTMePP porphyrinTTMePP porphyrin
H2TTMePP + H
2SO
4 [H2O]
[nm]360 380 400 420 440 460 480 500
A
0,00
0,20
0,40
0,60
0,80
1,00
1,20
H2P + 2 H+ = H4P2+
413 nm
436 nm
H2TTMePP + H
2SO
4 [H2O]
[nm]360 380 400 420 440 460 480 500
A
0,00
0,20
0,40
0,60
0,80
1,00
1,20
H2P + 2 H+ = H4P2+
413 nm
436 nm
dication:
H4P2+ H3P
+ + H+ (K4) monocation:
H3P+ H2P + H+ (K3)
free porphyrin:
H2P HP- + H+ (K2) monoanion:
HP- P2- + H+ (K1)
Four porphyrin acid-base equilibriaFour porphyrin acid-base equilibria
M-P
(H2P)2 (H2P)n
M H2P
(H4P2+ H3P
+ H2P HP-)
M-N H2P-N
N
where: H2P – free porphyrins,M – metals,N – nucleic bases, nucleosides and nucleotides.
Scheme of interactions between porphyrins systemsScheme of interactions between porphyrins systems and nucleic bases and their derivativesand nucleic bases and their derivatives
Nucleic bases and their derivativesNucleic bases and their derivatives
adenineadenine
adenosineadenosine
ATPATP
guanineguanine
guanosineguanosine
GTPGTP
Nucleic bases and their derivativesNucleic bases and their derivatives
thyminethymineuraciluracil
uridineuridine
UTPUTP CTPCTP
cytosinecytosine
cytidinecytidinethymidinethymidine
The sites of metal fixation in nucleic basesThe sites of metal fixation in nucleic bases
adenineadenine guanineguanine
thyminethyminecytosinecytosine
uraciluracil
nL
2L P(N)...P(N) PN N+P
N][]P(N)[]P(N)[
K1-n
nn
]P[[N]...KKK...[N]KK+[N]K1
[N]...KKK...]N[KK]N[K=A n
n212
211
nn21n
2212110
Association constantsAssociation constants
A - absorbancy;A - absorbancy;εε0 0 - molar absorbancy index of initial porphyrin concentration;- molar absorbancy index of initial porphyrin concentration;
εε11 and K and K11, ..., etc. - molar absorbancy indexes and gradual stability constants, ..., etc. - molar absorbancy indexes and gradual stability constants
for complexes with stoichiometry 1:1, 1:2, ..., etc;for complexes with stoichiometry 1:1, 1:2, ..., etc;[N] - molar nucleic base (ligand) concentration;[N] - molar nucleic base (ligand) concentration;[P] - molar porphyrin concentration.[P] - molar porphyrin concentration.
Titration experiments (pH=12.4)Titration experiments (pH=12.4)
H2TTMePP [Tris 0.025 M, pH =12.4] + Tris
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.0
0.1
0.20.3
0.4
0.50.6
0.70.8
0.9
1.0
1.1
1.21.3
1.4
1.5
1.6
1.7
1.8
Q band10 x414
421
516Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.4] + Tris
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.0
0.1
0.20.3
0.4
0.50.6
0.70.8
0.9
1.0
1.1
1.21.3
1.4
1.5
1.6
1.7
1.8
Q band10 x414
421
516Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.4] + uracil
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
Q band10 x
414
422
516
Soret band 560
H2TTMePP [Tris 0.025 M, pH =12.4] + uracil
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
Q band10 x
414
422
516
Soret band 560
H2TTMePP [Tris 0.025 M, pH =12.4] + uridine
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
Q band10 x
414
422
516Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.4] + uridine
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
Q band10 x
414
422
516Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.4] + UTP
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
Q band10 x
414
422
516
Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.4] + UTP
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
Q band10 x
414
422
516
Soret band
560
Titration experiments (pH=12.7)Titration experiments (pH=12.7)
H2TTMePP [Tris 0.025 M, pH =12.69] + Uracil
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.00.10.20.30.40.50.60.70.80.91.01.11.21.31.41.51.61.71.81.92.0
Q band10 x
414
422
516
Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.69] + Uracil
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.00.10.20.30.40.50.60.70.80.91.01.11.21.31.41.51.61.71.81.92.0
Q band10 x
414
422
516
Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.69] + Uridine
[nm]
350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.00.10.20.30.40.50.60.70.80.91.01.11.21.31.41.51.61.71.81.92.0
Q band10 x
414
422
516
Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.69] + Uridine
[nm]
350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0.00.10.20.30.40.50.60.70.80.91.01.11.21.31.41.51.61.71.81.92.0
Q band10 x
414
422
516
Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.69] + UTP
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
-0.10.00.10.20.30.40.50.60.70.80.91.01.11.21.31.41.51.61.71.81.92.0
Q band10 x
414
422
516Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.69] + UTP
[nm]350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
-0.10.00.10.20.30.40.50.60.70.80.91.01.11.21.31.41.51.61.71.81.92.0
Q band10 x
414
422
516Soret band
560
H2TTMePP [Tris 0.025 M, pH =12.69] + Tris
[nm]
350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0,00,10,20,30,40,50,60,70,80,91,01,11,21,31,41,51,61,71,8
Q band10 x
414
422
516Soret band 560
H2TTMePP [Tris 0.025 M, pH =12.69] + Tris
[nm]
350 375 400 425 450 475 500 525 550 575 600 625 650 675 700
A
0,00,10,20,30,40,50,60,70,80,91,01,11,21,31,41,51,61,71,8
Q band10 x
414
422
516Soret band 560
The absorbance changes The absorbance changes in the functionin the function
of porphyrin concentrationof porphyrin concentration
cM H2TTMePP * 1062.5 3.0 3.5 4.0
A
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
H2TTMePP - normalized plotpH = 12.7+ tris+ uracil+ uridine+ UTP
cM H2TTMePP * 1062.5 3.0 3.5 4.0
A
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
H2TTMePP - normalized plotpH = 12.7+ tris+ uracil+ uridine+ UTP
cM H2TTMePP * 1062.5 3.0 3.5 4.0 4.5
A
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
H2TTMePP - normalized plotpH = 12.4+ tris+ uracil+ uridine+ UTP
cM H2TTMePP * 1062.5 3.0 3.5 4.0 4.5
A
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
H2TTMePP - normalized plotpH = 12.4+ tris+ uracil+ uridine+ UTP
Association constantsAssociation constants
H2TMePyP CuTMePyP H2TTMePP CuTTMePP
pH=9.07 (titrant cM = 10-3)
adenine 7 221 29 1 226 16 19 054 28 6 734 10
adenosine 5 271 11 1 281 6 60 240 24 2 690 18
di-Na-ATP 6 912 5 9 346 4 224 460 25 32 440 18
pH=13.6 (titrant cM = 10-1)
adenine 1.89 3 2 077 15 285 6 129 5
adenosine 17.30 4 15 120 4 535 8 736 8
di-Na-ATP 13.09 3 12 125 8 4 250 9 3 119 9
pH=12.4 (titrant cM = 10-3)
guanine 57 884 2 26 326 20 177 500
7590 18 029 14
guanosine 2 581 4 4 255 25 147 500
3460 27 980 21
tri-Na-GTP 1 746 8 25 152 13 22 320 460 41 810 7
pH=13.0 (titrant cM = 10-2)
guanine 10-6 347 26 23 630 640 1 232 22
guanosine 7 8 1 109 8 21 620 840 643 4
H2TMePyP CuTMePyP H2TTMePP CuTTMePP
pH=9.07 (titrant cM = 10-3)
adenine 7 221 29 1 226 16 19 054 28 6 734 10
adenosine 5 271 11 1 281 6 60 240 24 2 690 18
di-Na-ATP 6 912 5 9 346 4 224 460 25 32 440 18
pH=13.6 (titrant cM = 10-1)
adenine 1.89 3 2 077 15 285 6 129 5
adenosine 17.30 4 15 120 4 535 8 736 8
di-Na-ATP 13.09 3 12 125 8 4 250 9 3 119 9
pH=12.4 (titrant cM = 10-3)
guanine 57 884 2 26 326 20 177 500
7590 18 029 14
guanosine 2 581 4 4 255 25 147 500
3460 27 980 21
tri-Na-GTP 1 746 8 25 152 13 22 320 460 41 810 7
pH=13.0 (titrant cM = 10-2)
guanine 10-6 347 26 23 630 640 1 232 22
guanosine 7 8 1 109 8 21 620 840 643 4
H2TMePyP CuTMePyP H2TTMePP CuTTMePP
pH=12.4 (titrant cM = 10-3)
uracil 1 890 5 5 354 38 95 250 21 9 407 30
uridine 3 756 23 23 733 27 10 989 33 30 123 14
tri-Na-UTP 5 265 4 60 078 20 71 475 16 57 123 13
pH=12.7 (titrant cM = 10-2)
uracil 441 5 1 634 30 10 935 14 3 363 17
uridine 6 083 6 10 195 8 3 021 20 1 191 23
tri-Na-UTP 6 776 14 46 410 11 5 538 20 4 021 13
pH=12.1 (titrant cM = 10-3)
thymine 1 289 4 10 111 4 32 527 23 5 967 26
thymidine 3 330 2 19 896 3 9 078 3 10 207 3
pH=12.1 (titrant cM = 10-2)
thymine 811 2 1 226 20 1 582 4 2 055 3
thymidine 554 2 3 614 19 2 142 19 1 043 4
pH=9.3 (titrant cM = 10-3)
cytosine 15 128 2 1 680 5 32 340 8 59 733 15
cytidine 4 141 4 4 670 4 21 214 13 22 399 3
tri-Na-CTP 8 326 2 5 418 3 60 664 10 13 149 2
pH=9.4 (titrant cM = 10-2)
cytosine 849 2 1 229 2 771 2 4 933 12
cytidine 3 716 3 5 124 1 1 042 3 605 3
tri-Na-CTP 1 543 6 9 389 6 6 873 11 1 863 13
H2TMePyP CuTMePyP H2TTMePP CuTTMePP
pH=12.4 (titrant cM = 10-3)
uracil 1 890 5 5 354 38 95 250 21 9 407 30
uridine 3 756 23 23 733 27 10 989 33 30 123 14
tri-Na-UTP 5 265 4 60 078 20 71 475 16 57 123 13
pH=12.7 (titrant cM = 10-2)
uracil 441 5 1 634 30 10 935 14 3 363 17
uridine 6 083 6 10 195 8 3 021 20 1 191 23
tri-Na-UTP 6 776 14 46 410 11 5 538 20 4 021 13
pH=12.1 (titrant cM = 10-3)
thymine 1 289 4 10 111 4 32 527 23 5 967 26
thymidine 3 330 2 19 896 3 9 078 3 10 207 3
pH=12.1 (titrant cM = 10-2)
thymine 811 2 1 226 20 1 582 4 2 055 3
thymidine 554 2 3 614 19 2 142 19 1 043 4
pH=9.3 (titrant cM = 10-3)
cytosine 15 128 2 1 680 5 32 340 8 59 733 15
cytidine 4 141 4 4 670 4 21 214 13 22 399 3
tri-Na-CTP 8 326 2 5 418 3 60 664 10 13 149 2
pH=9.4 (titrant cM = 10-2)
cytosine 849 2 1 229 2 771 2 4 933 12
cytidine 3 716 3 5 124 1 1 042 3 605 3
tri-Na-CTP 1 543 6 9 389 6 6 873 11 1 863 13
ConclusionsConclusions11. The big differences in the results for particular nucleic agents, and first of all for different pH . The big differences in the results for particular nucleic agents, and first of all for different pH values indicate the diversification ofvalues indicate the diversification of interaction level of investigated systems. Such results can be interaction level of investigated systems. Such results can be
the proof ofmutualthe proof ofmutual interactions porphyrin - nucleic ligand.interactions porphyrin - nucleic ligand.
2. Interactions of H2. Interactions of H22TTMePP with nucleic bases are much stronger than interactions of TTMePP with nucleic bases are much stronger than interactions of
HH22TMePyP. Such effect is caused by the aniline group, much bigger than pyridyl group - TMePyP. Such effect is caused by the aniline group, much bigger than pyridyl group -
“stacking” (specific interactions consisting in forming of associates by reacting molecules) between “stacking” (specific interactions consisting in forming of associates by reacting molecules) between porphyrin and nucleic ligands is much more stronger for bigger compounds.porphyrin and nucleic ligands is much more stronger for bigger compounds.
3. The results show that the more stable associates are formed at lower pH value. Moreover, the 3. The results show that the more stable associates are formed at lower pH value. Moreover, the strength ofstrength of the obtained associates increases in series:nucleic base the obtained associates increases in series:nucleic base nucleoside nucleoside nucleotide, nucleotide,
although many departures from the rule are observed, because of high diversity and different pH although many departures from the rule are observed, because of high diversity and different pH values of investigatedvalues of investigated systems.systems.
4. The association constants for interactions between porphyrins and nucleic agents let us suppose 4. The association constants for interactions between porphyrins and nucleic agents let us suppose that by means of adequate porphyrin substituent groups and metal ion in porphyrin cave it would that by means of adequate porphyrin substituent groups and metal ion in porphyrin cave it would be possible to control the level of interactions between porphyrins and complicated DNA systems be possible to control the level of interactions between porphyrins and complicated DNA systems
and, probabely, introduce metalloporphyrins to the desirable place of DNA chain.and, probabely, introduce metalloporphyrins to the desirable place of DNA chain.
5. However, the substitution reactions proceeding on the metallic centres connected with nucleic 5. However, the substitution reactions proceeding on the metallic centres connected with nucleic acids are much more complicated, what is caused by the interactions of ligands with other groups, acids are much more complicated, what is caused by the interactions of ligands with other groups, as well as conformational changes in a macromolecule. Therefore all data obtained during study of as well as conformational changes in a macromolecule. Therefore all data obtained during study of porphyrin - DNA interactions should be particularly considered, because some porphyrin systems porphyrin - DNA interactions should be particularly considered, because some porphyrin systems
can join and interact with DNA in different ways, depending on neighbouring chemical can join and interact with DNA in different ways, depending on neighbouring chemical compounds. Very often the manner of interactions between reagents is determined by the kind of compounds. Very often the manner of interactions between reagents is determined by the kind of
reaction buffer and ionic strength connected with environment of investigated system.reaction buffer and ionic strength connected with environment of investigated system.
Metalloporphyrin molecule incorporated in DNA chainMetalloporphyrin molecule incorporated in DNA chain