Abstract
Phenylboronic acid groups as sugar recognition sites were successfully introduced into native myoglobin by a cofactor-reconstitution method. Spectrophotometric pH titration demonstrated the sugar-induced pK(a) shift of the H2O coordinated to the heme center of the semisynthetic myoglobin bearing phenylboronic acids (met-Mb(PhBOH)2). By means of circular dichroism (CD) and paramagnetic 1H NMR spectroscopies, it was proven that sugars that were bound to phenylboronic acid sites induced the rearrangement of the heme crevice to reinforce the heme cofactor-apoprotein interactions. The structural changes that were induced by the binding of sugars subsequently enhanced the dioxygen storage activity of Mb(PhBOH)2. Such sugar-induced structural and functional changes did not occur for other modified Mbs that had no sugar-recognition units. Interestingly, a randomly modified Mb with phenylboronic acid units did not show any sugar response. In Mb(PhBOH)2, the information from the sugar-binding event was efficiently transmitted to the active center, so that the activity was efficiently altered upon sugar binding. In conclusion, the active site specific incorporation of molecular recognition units as nonnatural functional molecules can provide a novel strategy for the design of stimuli-responsive semisynthetic proteins.
| Original language | English |
|---|---|
| Pages (from-to) | 1025-1031 |
| Number of pages | 7 |
| Journal | Chemistry - A European Journal |
| Volume | 3 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - Jan 1 1997 |
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All Science Journal Classification (ASJC) codes
- Catalysis
- Chemistry(all)
- Organic Chemistry
Cite this
Design and synthesis of sugar-responsive semiartificial myoglobin triggered by modulation of apoprotein-cofactor interactions. / Hamachi, Itaru; Tajiri, Yusuke; Nagase, Tsuyoshi; Shinkai, Seiji.
In: Chemistry - A European Journal, Vol. 3, No. 7, 01.01.1997, p. 1025-1031.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Design and synthesis of sugar-responsive semiartificial myoglobin triggered by modulation of apoprotein-cofactor interactions
AU - Hamachi, Itaru
AU - Tajiri, Yusuke
AU - Nagase, Tsuyoshi
AU - Shinkai, Seiji
PY - 1997/1/1
Y1 - 1997/1/1
N2 - Phenylboronic acid groups as sugar recognition sites were successfully introduced into native myoglobin by a cofactor-reconstitution method. Spectrophotometric pH titration demonstrated the sugar-induced pK(a) shift of the H2O coordinated to the heme center of the semisynthetic myoglobin bearing phenylboronic acids (met-Mb(PhBOH)2). By means of circular dichroism (CD) and paramagnetic 1H NMR spectroscopies, it was proven that sugars that were bound to phenylboronic acid sites induced the rearrangement of the heme crevice to reinforce the heme cofactor-apoprotein interactions. The structural changes that were induced by the binding of sugars subsequently enhanced the dioxygen storage activity of Mb(PhBOH)2. Such sugar-induced structural and functional changes did not occur for other modified Mbs that had no sugar-recognition units. Interestingly, a randomly modified Mb with phenylboronic acid units did not show any sugar response. In Mb(PhBOH)2, the information from the sugar-binding event was efficiently transmitted to the active center, so that the activity was efficiently altered upon sugar binding. In conclusion, the active site specific incorporation of molecular recognition units as nonnatural functional molecules can provide a novel strategy for the design of stimuli-responsive semisynthetic proteins.
AB - Phenylboronic acid groups as sugar recognition sites were successfully introduced into native myoglobin by a cofactor-reconstitution method. Spectrophotometric pH titration demonstrated the sugar-induced pK(a) shift of the H2O coordinated to the heme center of the semisynthetic myoglobin bearing phenylboronic acids (met-Mb(PhBOH)2). By means of circular dichroism (CD) and paramagnetic 1H NMR spectroscopies, it was proven that sugars that were bound to phenylboronic acid sites induced the rearrangement of the heme crevice to reinforce the heme cofactor-apoprotein interactions. The structural changes that were induced by the binding of sugars subsequently enhanced the dioxygen storage activity of Mb(PhBOH)2. Such sugar-induced structural and functional changes did not occur for other modified Mbs that had no sugar-recognition units. Interestingly, a randomly modified Mb with phenylboronic acid units did not show any sugar response. In Mb(PhBOH)2, the information from the sugar-binding event was efficiently transmitted to the active center, so that the activity was efficiently altered upon sugar binding. In conclusion, the active site specific incorporation of molecular recognition units as nonnatural functional molecules can provide a novel strategy for the design of stimuli-responsive semisynthetic proteins.
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U2 - 10.1002/chem.19970030707
DO - 10.1002/chem.19970030707
M3 - Article
AN - SCOPUS:0030842481
VL - 3
SP - 1025
EP - 1031
JO - Chemistry - A European Journal
JF - Chemistry - A European Journal
SN - 0947-6539
IS - 7
ER -