Supporting Information
Development of Bifunctional Photoactivatable
Benzophenone Probes and their Application to
Glycoside Substrates
Nir Qvit1,*, Galya Monderer-Rothkof2, Ayelet Ido2, Deborah E. Shalev3, Orna AmsterChoder2 and Chaim Gilon1,*
1
Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel and
2
Department of Molecular Biology, The Hebrew University Medical School, Jerusalem,
91120, Israel, and 3The Wolfson Centre for Applied Structural Biology, The Hebrew
University of Jerusalem, Jerusalem, 91904, Israel.
*To whom correspondence should be addressed. Institute of Chemistry, The Hebrew
University of Jerusalem, Jerusalem 91904, Israel. Phone: +972-2-6586181. Fax: +972-26515731. Email: nir.qvit@gmail.com, gilon@vms.huji.ac.il.
Contents
Structure and characterization of 7
Pages
S2 – S6
Structure and characterization of 4
S7 – S9
Structure and characterization of 5
S10 – S11
Structure and characterization of 6
S12 – S14
S1
Figure S1. Structure and characterization of 7.
Structure (A) and MS (B) of 7. The peaks in the MS are 481.2 m/z, MW of 7 + H, and 319.1
m/z MW of the fragment of 7 (marked in Figure A, in the box) obtained from the MS
analysis, (C) Analytical HPLC of product 7, (D) 1H NMR of 7 in ACN-d3, (E) 13C NMR of 7
in ACN-d3, (F) COSY spectrum of 7 in ultra-dry ACN-d3 showing aromatic (left) and
aliphatic (right) regions of spectrum, (G) ROESY spectrum of 7 in ultra-dry ACN-d3 showing
specific interactions designated in text, which give the connectivity of the molecule.
S2
S3
F
S4
G
Proton
Proton chemical shift (ppm), multiplicity (d-doublet, t-triplet, mDesignation multiplet), (integration), J-coupling (Hz), 600 MHz spectrometer.
H1
H 4.98 ppm, d (1H)
H2
H 3.47 ppm, unresolved
OH2
H 3.75 ppm, d (1H), J=3.3 Hz
H3
H 3.47 ppm, unresolved
OH3
H 3.59 ppm, d (1H), J=2.6 Hz
H4
H 3.44 ppm, m (1H)
OH4
H 3.47 ppm, unresolved
H5
H 3.51 ppm, m (1H)
CH26
H 3.87, 3.70 ppm, m (2H)
OH6
H 2.92 ppm, t (1H), J=6.2 Hz
H12
H 7.74 ppm, s, broad (1H)
H8
H 7.28 ppm, dd (1H) J=9.0, 3.0
H9
H 7.07 ppm, d (1H) J=9.0 Hz
OH10
H 11.80 ppm, s (b, 1H)
H14
H 8.11 ppm, dm (2H) J=8.6, 1.9 Hz
S5
Proton
Proton chemical shift (ppm), multiplicity (d-doublet, t-triplet, mDesignation multiplet), (integration), J-coupling (Hz), 600 MHz spectrometer.
H15
H 8.01 ppm, dm (2H) J=8.6, 1.9 Hz
H19
H 7.89 ppm, dm (2H) J=8.4 Hz
H20
H 7.62 ppm, tm (2H) unresolved
H21
H 7.75 ppm, tt (b, 1H) J=7.5, 1.3 Hz
Assignment considerations:
Chemical shift considerations determined that H1 was the low-field signal in the aliphatic
region. The H1 peak showed a single interaction with H2, which showed a strong interaction
with its hydroxyl, OH2 and a weak interaction with OH3. All five hydroxy groups in the
molecule showed exchange peaks with water in the ROESY spectrum. The OH6 peak was
the only OH peak with triplet multiplicity. It showed COSY interactions with both C6
methylene protons, which both interacted with H5 and each other. H5 interacted with H4. H3
and OH4 are assigned as part of the unresolved multiplet at H 3.47 ppm, which also shows
an exchange peak with water in the ROESY spectrum.
The aromatic spin systems comprising protons H14-H15 and H19-H20-H21 were assigned
according to the number of spins in the system, their connectivity and multiplicity. H14 and
H15 showed COSY interactions between them, and H14 showed a ROESY interaction with
OH10 whereas H15 did not, thereby giving the directionality of this system. The H19-H20H12 protons were in a unique system with unambiguous coupling and integration values.
The aromatic spin system comprising H8-H9-OH10-H12 showed COSY interactions between
H8 and both H9 and H12. Coupling values gave the assignment; however the specific
connectivity of the azo bond was not evident from the correlation spectrum, and is discussed
in the text.
S6
Figure S2. Structure and characterization of 4.
(A) Structure of 4, (B) MS of 4. The peaks in the MS are: 538.2 m/z – MW of 4 + H, 560.2
m/z - MW of 4 + Na and 376.1 m/z The fragment of 4 (marked in Figure A, in the box)
obtained from the MS analysis, (C) Analytical HPLC of 4, (D) 1H NMR of 4 in ACN-d3, (E)
1
H NMR of 4 in DMSO-d6. (F) 13C NMR of 4 in DMSO-d6.
S7
S8
S9
Figure S3. Structure and characterization of 5.
(A) Structure of 5, (B) MS of 5. The peaks in the MS are: 647.4 m/z – MW of 5 + H,
669.4 m/z - MW of 5 + Na and 485.3 m/z, the fragment of 5 (marked in Figure A, in
the box) obtained from the MS analysis, (C) Analytical HPLC of 5, (D) 1H NMR of 5
in ACN-d3, and (E) 13C NMR of 5 in ACN-d3.
S10
S11
Figure S4. Structure and characterization of 6.
(A) Structure of 6, (B) MS of 6. The peaks in the MS are: 793.2 m/z – MW of 6 + H,
and 630.2 m/z, the fragment of 6 (marked in Figure A, in the box) obtained from the
MS analysis, (C) Analytical HPLC of 6, (D) 1H NMR of 6 in DMSO-d6 at 25 °C, (E)
1
H NMR of 6 in DMSO-d6 at 45 °C and (F) 13C NMR of 6 in DMSO-d6.
S12
S13
S14
S15