Ultrafast Proton Coupled Electron Transfer between Tryptophan and Tyrosine in Peptides Trp-Pro_n-Tyr

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State Key Laboratory of Precision Spectroscopy, East China Normal University, Shanghai 200241, China
2.
Key Laboratory of 3D Micro/Nano Fabrication and Characterization of Zhejiang Province, School of Engineering, Westlake University, Hangzhou 310024, China

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Corresponding author: E-mail: jqchen@lps.ecnu.edu.cn; jhxu@phy.ecnu.edu.cn

摘要

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Abstract: A series of model peptides (Trp–Pro_n–Tyr, WP_nY, n=0, 1, 2, 3, 5), which contain tryptophan (Trp, W), tyrosine (Tyr, Y), and proline (Pro, P), have been studied under three typical pH conditions (3, 7, and 10) by steady-state absorption and fluorescence spectroscopy, nanosecond time-resolved fluorescence and femtosecond time-resolved transient absorption spectroscopy. When the peptide’s chain length is increased, Trp fluorescence quenching is expected to be gradually weakened. However, Trp fluorescence in WPY is strongly quenched and reveals even stronger quenching with increasing pH values, whose hypochromicity is clearly different from other model peptides. Transient absorption spectra also demonstrate that the excited state decay of WPY is much faster than that of other model peptides, especially at pH = 10. It is attributed to the efficient proton coupled electron transfer (PCET) between Trp and Tyr. Moreover, due to the very short distance between Trp and Tyr in WPY, this PCET process could be achieved by “direct transfer”, contrasted with the slow and long-range PCET process in other model peptides. Our results of the dipeptides WY and WP further suggest that Trp may also have more complex interactions with the peptide backbone or proline in those peptides. This work provides an experimental evidence for the electron transfer mechanism in WY dyads, which can help ones to understand how to reduce Trp radicals in proteins.
- Proton coupled electron transfer /
- Tryptophan /
- Tyrosine /
- Peptides /
- Spectroscopy /
- Dynamics

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Figure 1. Normalized absorption spectra (A, B, and C) and fluorescence spectra (D, E, and F) of W, WP, and WP_nY (n=0, 1, 2, 3, 5) at three typical pH values (pH=3, 7, and 10). The excitation wavelength in the fluorescence spectra (D–F) is 295 nm.

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Figure 2. Normalized nanosecond time-resolved fluorescence decay curves of W, WP, and WP_nY (n=0, 1, 2, 3, 5) with pH=3 (A), 7 (B), and 10 (C). All fluorescence decay curves are excited at 295 nm.

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Figure 3. Femtosecond TA spectra of W (A), WY (B), WP (C), WPY (D), WP₂Y (E), WP₃Y (F), and WP₅Y (G) at pH=3 with the excitation wavelength at 295 nm, respectively. Time window is from 2 ps to 7 ns.

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Figure 4. Femtosecond TA spectra of W (A), WY (B), WP (C), WPY (D), WP₂Y (E), WP₃Y (F), and WP₅Y (G) at pH=7 with the excitation wavelength at 295 nm, respectively. Time window is from 2 ps to 7 ns.

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Figure 5. Femtosecond TA spectra of W (A), WY (B), WP (C), WPY (D), WP₂Y (E), WP₃Y (F), and WP₅Y (G) at pH=10 with the excitation wavelength at 295 nm, respectively. Time window is from 2 ps to 7 ns.

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Figure 6. Normalized femtosecond TA kinetics (at 580 nm) of W, WP and WP_nY (n=0, 1, 2, 3, 5) with pH=3 (A), 7 (B) and 10 (C), respectively. The data collected by the TA system are represented by circles, and the fitting results are represented by lines.

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Table I. The lifetimes ($ {\tau }_{n} $ in unit of ns) and amplitudes ($A_{n}$ in unit of %) of W, WP, and WP_nY (n =0, 1, 2, 3, 5) by fitting the decay curves obtained from TCSPC. The average lifetime (om TCSPC. The average lifetime (${\tau }_{\rm{ave}}$ in unit of ns) was also calculated.

Peptide	pH=3					pH=7							pH=10
	$A_{1}$	$ {\tau }_{1} $	$A_{2}$	$ {\tau }_{2} $	${\tau }_{\rm{ave}}$	$A_{1}$	$ {\tau }_{1} $	$A_{2}$	$ {\tau }_{2} $	$A_{3}$	$ {\tau }_{3} $	${\tau }_{\rm{ave}}$	$A_{1}$	$ {\tau }_{1} $	$A_{2}$	$ {\tau }_{2} $	$A_{3}$	$ {\tau }_{3} $	${\tau }_{\rm{ave}}$
W	28	0.45	72	2.71	2.57	5	0.40	95	2.90	–	–	2.88	42	2.49	58	8.28	–	–	7.24
WY	44	0.59	56	1.69	1.45	29	0.29	50	1.61	21	4.93	3.31	11	0.36	19	1.74	70	4.97	4.64
WP	68	0.40	32	2.01	1.53	61	0.35	29	2.25	10	6.06	3.50	11	0.59	69	4.32	20	7.53	5.32
WPY	61	0.32	39	1.73	1.40	56	0.22	32	1.35	12	3.27	2.00	56	0.20	25	1.42	19	3.36	2.42
WP₂Y	53	0.31	47	2.05	1.80	50	0.23	42	2.03	8	5.82	3.10	23	0.52	44	2.32	33	5.70	4.36
WP₃Y	52	0.30	48	2.06	1.82	55	0.24	39	2.10	6	6.80	3.33	16	0.37	46	2.24	38	6.43	5.09
WP₅Y	55	0.27	45	2.16	1.90	52	0.22	41	2.11	7	6.97	3.56	11	0.49	49	2.31	40	7.21	5.77

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Table II. The lifetimes ($ {\tau }_{n} $ in unit of ns) and amplitudes ($A_{n}$ in unit of %) by fitting the decay curves obtained in FIG. 6.

Peptide	pH=3				pH=7				pH=10
	$A_{1}$	$ {\tau }_{1} $	$A_{2}$	$ {\tau }_{2} $	$A_{1}$	$ {\tau }_{1} $	$A_{2}$	$ {\tau }_{2} $	$A_{1}$	$ {\tau }_{1} $	$A_{2}$	$ {\tau }_{2} $
W	43	1.03	57	8.43	50	2.57	50	6.98	100	12.30	–	–
WY	56	0.57	44	7.35	50	0.96	50	6.74	11	0.12	89	10.92
WP	64	0.30	36	4.94	69	0.37	31	6.54	19	0.11	81	11.87
WPY	58	0.126	42	3.22	64	0.15	36	2.59	59	0.08	41	9.54
WP₂Y	58	0.17	42	3.89	57	0.29	43	4.18	35	0.14	65	9.76
WP₃Y	50	0.23	50	4.23	54	0.25	46	5.00	37	0.12	63	11.13
WP₅Y	59	0.27	41	5.08	58	0.23	42	5.05	33	0.15	67	10.06

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