Steady-state and time-resolved fluorescence spectroscopic studies on interaction of the N-terminal region with the hairpin loop of the phytocystain Scb

Keiko Doi-Kawano, Etsuko Nishimoto, Yoshiaki Kouzuma, Daisuke Takahashi, Shoji Yamashita, Makoto Kimura

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The steady-state and time-resolved fluorescece spectroscopy is one of the most powerful method to detect and analyze subtle conformation change and interaction between peptide elements in protein. Phytocystatin Scb isolated from sunflower seeds includes a single Trp residue at position 85. In an attempt to investigate the interaction of the N-terminal region of Scb with the first and second hairpin loops by fluorescence spectroscopy of Trp residue, two Scb mutants in which single Trp locates at position 52 and 58, respectively, and their N-terminal removed mutants were generated. The N-terminal truncation changed the fluorescence decay kinetics of Trp52 from the triple exponential to double. Furthermore, the time-resolved fluorescence anisotropy residue indicated that the segmental motion of Trp52 was significantly enhanced by its N-terminal truncation. In contrast, Trp58 and Trp85 had little influence. The N-terminal successive truncations of Scb and its mutants resulted in the weaken inhibitors to papain. These results suggested that the N-terminal region of Scb interacts with the peptide segment preceding the first hairpin loop, thereby stabilizing the conformation of the hairpin loop structure.

Original languageEnglish
Pages (from-to)631-639
Number of pages9
JournalJournal of Fluorescence
Volume19
Issue number4
DOIs
Publication statusPublished - Jul 1 2009

Fingerprint

Conformations
Fluorescence
Peptides
Fluorescence Polarization
Papain
Helianthus
Fluorescence Spectrometry
Fluorescence spectroscopy
interaction
Seed
Spectrum Analysis
Seeds
Anisotropy
Spectroscopy
Kinetics
Proteins
time

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Clinical Psychology
  • Social Sciences (miscellaneous)
  • Sociology and Political Science
  • Spectroscopy
  • Clinical Biochemistry
  • Law

Cite this

Steady-state and time-resolved fluorescence spectroscopic studies on interaction of the N-terminal region with the hairpin loop of the phytocystain Scb. / Doi-Kawano, Keiko; Nishimoto, Etsuko; Kouzuma, Yoshiaki; Takahashi, Daisuke; Yamashita, Shoji; Kimura, Makoto.

In: Journal of Fluorescence, Vol. 19, No. 4, 01.07.2009, p. 631-639.

Research output: Contribution to journalArticle

@article{c528c3a9c57a41128b306079b31abf69,
title = "Steady-state and time-resolved fluorescence spectroscopic studies on interaction of the N-terminal region with the hairpin loop of the phytocystain Scb",
abstract = "The steady-state and time-resolved fluorescece spectroscopy is one of the most powerful method to detect and analyze subtle conformation change and interaction between peptide elements in protein. Phytocystatin Scb isolated from sunflower seeds includes a single Trp residue at position 85. In an attempt to investigate the interaction of the N-terminal region of Scb with the first and second hairpin loops by fluorescence spectroscopy of Trp residue, two Scb mutants in which single Trp locates at position 52 and 58, respectively, and their N-terminal removed mutants were generated. The N-terminal truncation changed the fluorescence decay kinetics of Trp52 from the triple exponential to double. Furthermore, the time-resolved fluorescence anisotropy residue indicated that the segmental motion of Trp52 was significantly enhanced by its N-terminal truncation. In contrast, Trp58 and Trp85 had little influence. The N-terminal successive truncations of Scb and its mutants resulted in the weaken inhibitors to papain. These results suggested that the N-terminal region of Scb interacts with the peptide segment preceding the first hairpin loop, thereby stabilizing the conformation of the hairpin loop structure.",
author = "Keiko Doi-Kawano and Etsuko Nishimoto and Yoshiaki Kouzuma and Daisuke Takahashi and Shoji Yamashita and Makoto Kimura",
year = "2009",
month = "7",
day = "1",
doi = "10.1007/s10895-008-0454-7",
language = "English",
volume = "19",
pages = "631--639",
journal = "Journal of Fluorescence",
issn = "1053-0509",
publisher = "Springer New York",
number = "4",

}

TY - JOUR

T1 - Steady-state and time-resolved fluorescence spectroscopic studies on interaction of the N-terminal region with the hairpin loop of the phytocystain Scb

AU - Doi-Kawano, Keiko

AU - Nishimoto, Etsuko

AU - Kouzuma, Yoshiaki

AU - Takahashi, Daisuke

AU - Yamashita, Shoji

AU - Kimura, Makoto

PY - 2009/7/1

Y1 - 2009/7/1

N2 - The steady-state and time-resolved fluorescece spectroscopy is one of the most powerful method to detect and analyze subtle conformation change and interaction between peptide elements in protein. Phytocystatin Scb isolated from sunflower seeds includes a single Trp residue at position 85. In an attempt to investigate the interaction of the N-terminal region of Scb with the first and second hairpin loops by fluorescence spectroscopy of Trp residue, two Scb mutants in which single Trp locates at position 52 and 58, respectively, and their N-terminal removed mutants were generated. The N-terminal truncation changed the fluorescence decay kinetics of Trp52 from the triple exponential to double. Furthermore, the time-resolved fluorescence anisotropy residue indicated that the segmental motion of Trp52 was significantly enhanced by its N-terminal truncation. In contrast, Trp58 and Trp85 had little influence. The N-terminal successive truncations of Scb and its mutants resulted in the weaken inhibitors to papain. These results suggested that the N-terminal region of Scb interacts with the peptide segment preceding the first hairpin loop, thereby stabilizing the conformation of the hairpin loop structure.

AB - The steady-state and time-resolved fluorescece spectroscopy is one of the most powerful method to detect and analyze subtle conformation change and interaction between peptide elements in protein. Phytocystatin Scb isolated from sunflower seeds includes a single Trp residue at position 85. In an attempt to investigate the interaction of the N-terminal region of Scb with the first and second hairpin loops by fluorescence spectroscopy of Trp residue, two Scb mutants in which single Trp locates at position 52 and 58, respectively, and their N-terminal removed mutants were generated. The N-terminal truncation changed the fluorescence decay kinetics of Trp52 from the triple exponential to double. Furthermore, the time-resolved fluorescence anisotropy residue indicated that the segmental motion of Trp52 was significantly enhanced by its N-terminal truncation. In contrast, Trp58 and Trp85 had little influence. The N-terminal successive truncations of Scb and its mutants resulted in the weaken inhibitors to papain. These results suggested that the N-terminal region of Scb interacts with the peptide segment preceding the first hairpin loop, thereby stabilizing the conformation of the hairpin loop structure.

UR - http://www.scopus.com/inward/record.url?scp=70349261441&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70349261441&partnerID=8YFLogxK

U2 - 10.1007/s10895-008-0454-7

DO - 10.1007/s10895-008-0454-7

M3 - Article

C2 - 19104918

AN - SCOPUS:70349261441

VL - 19

SP - 631

EP - 639

JO - Journal of Fluorescence

JF - Journal of Fluorescence

SN - 1053-0509

IS - 4

ER -