I don't have a definitive answer for you, but you may glean the information
you need by looking at Aebersold's papers on the use of a novel
thiohydantoin, synthesised to give good coupling characteristics and high
detection sensitvity. Some abstracts are below.
Len
TI: High-sensitivity detection of 4-(3-pyridinylmethylaminocarboxypropyl)
phenylthiohydantoins by capillary liquid chromatography
microelectrospray ion trap mass spectrometry
AU: Ducret_A, Bures_EJ, Aebersold_R
NA: UNIV WASHINGTON,DEPT MOL BIOTECHNOL,SEATTLE,WA,98195
UNIV WASHINGTON,DEPT MOL BIOTECHNOL,SEATTLE,WA,98195
AMGEN INC,THOUSAND OAKS,CA,91320
JN: JOURNAL OF PROTEIN CHEMISTRY, 1997, Vol.16, No.5, pp.323-328
IS: 0277-8033
AB: We describe the separation and detection at the low-femtomole level
of 4-(3-pyridinylmethylaminocarboxypropyl) phenylthiohydantoins (311-
PTHs) by capillary liquid chromatography-microelectrospray ion trap
mass spectrometry. Highest sensitivity was obtained in the multiple-
ion monitoring operating mode in which we detected 311-PTHs at the 5-
fmol level with a signal-to-noise ratio of approximately 10. We
investigated the fragmentation patterns of the isobaric 311-PTH
isoleucine and 311-PTH leucine by electrospray ionization ion trap
tandem mass spectrometry. The compounds could be differentiated by a
fragment ion of mass m/z = 366.1 which was specific for the breakdown
of 311-PTH leucine, thus allowing for the unambiguous identification
of the 311-PTH derivatives of all 20 naturally occurring amino acids
by their masses and fragmentation patterns.
KP: PROTEIN
WA: protein sequencing, 311-PITC, capillary liquid chromatography,
microelectrospray, ion trap mass spectrometry
CR: AEBERSOLD_R, 1992 Vol.1 p.494, PROTEIN SCI
BASIC_C, 1995 Vol.6 p.1211, J AM SOC MASS SPECTR
BURES_EJ, 1995 Vol.224 p.363, ANAL BIOCHEM
EDMAN_P, 1949 Vol.22 p.475, ARCH BIOCHEM BIOPHYS
HESS_D, 1995 Vol.224 p.373, ANAL BIOCHEM
NIKA_H, 1994 Vol.13 p.439, J PROTEIN CHEM
STOLOWITZ_ML, 1993 p.37, METHODS PROTEIN SEQU
TI: LIQUID-CHROMATOGRAPHY ELECTROSPRAY-IONIZATION MASS-SPECTROMETRY OF 4-
(3-PYRIDINYLMETHYLAMINOCARBOXYPROPYL) PHENYLTHIOHYDANTOINS
AU: HESS_D, NIKA_H, CHOW_DT, BURES_EJ, MORRISON_HD, AEBERSOLD_R
NA: UNIV WASHINGTON,DEPT MOLEC BIOTECHNOL,FJ-20,SEATTLE,WA,98195
UNIV BRITISH COLUMBIA,BIOMED RES CTR,VANCOUVER,BC V6T 1Z3,CANADA
JN: ANALYTICAL BIOCHEMISTRY, 1995, Vol.224, No.1, pp.373-381
IS: 0003-2697
AB: We report the separation of 4-(3-pyridinylmethylaminocarboxypropyl)
phenylthiohydantoins by microbore reverse-phase high-performance
liquid chromatography and their detection by on-line electrospray
ionization mass spectrometry. These compounds are the products of the
chemical stepwise degradation of polypeptides using 4-(3-
pyridinylmethylaminocarboxypropyl) phenyl isothiocyanate. We describe
chromatographic conditions for on-column concentration of the
analytes and for baseline separation of the isobaric amino acid
derivatives of leucine and isoleucine. A commercially available
protein sequencer was readily interfaced with the described
analytical system and used for adsorptive sequence analysis of a
panel of synthetic peptides containing collectively all 20 naturally
occurring amino acids. On-line mass analysis of derivatives generated
by automated sequencing confirmed that the derivatives were of the
predicted mass and were detectable at comparable signal strength and
sensitivity. Finally, we demonstrate that the additional selectivity
in data interpretation provided by mass analysis dramatically
improves the signal-to-noise ratio and therefore enhances the ability
to conclusively interpret protein and peptide sequence data. (C) 1995
Academic Press,Inc.
TI: SYNTHESIS OF THE PROTEIN-SEQUENCING REAGENT 4-(3-
PYRIDINYLMETHYLAMINOCARBOXYPROPYL) PHENYL ISOTHIOCYANATE AND
CHARACTERIZATION OF 4-(3-PYRIDINYLMETHYLAMINOCARBOXYPROPYL)
PHENYLTHIOHYDANTOINS
AU: BURES_EJ, NIKA_H, CHOW_DT, MORRISON_HD, HESS_D, AEBERSOLD_R
NA: UNIV WASHINGTON,DEPT MOLEC BIOTECHNOL,FJ-20,SEATTLE,WA,98195
UNIV BRITISH COLUMBIA,BIOMED RES CTR,VANCOUVER,BC V6T 1Z3,CANADA
JN: ANALYTICAL BIOCHEMISTRY, 1995, Vol.224, No.1, pp.364-372
IS: 0003-2697
AB: We report the synthesis and structural characterization of the novel
Edman-type protein-sequencing reagent 4-(3-
pyridinylmethylaminocarboxypropyl) phenyl isothiocyanate. A panel of
thiohydantoins prepared from this reagent were found stable during
liquid chromatography-electrospray mass spectrometry and were
detectable at the low femtomole sensitivity level. Furthermore, the
signal detected for these compounds in the mass spectrometer was
linear from the low femtomole to the low picomole range. The
derivatives showed uv absorbance spectra comparable to their
phenylthiohydantoin counterparts. The extinction coefficient for the
4-(3-pyridinylmethylaminocarboxypropyl) phenyl thiohydantoin tyrosine
was determined by adsorptive sequence analysis of a synthetic
pentapeptide featuring an N-terminal I-125-labeled tyrosine. The
sequence data suggest that the reagent will be useful for extended
sequence analysis of proteins and peptides using commercially
available gas-liquid-phase sequencers. (C) 1995 Academic Press, Inc.
*********************************************************************
Dr Len C. Packman
Assistant Director of Research
Protein and Nucleic Acid Chemistry Facility
Department of Biochemistry
University of Cambridge
80 Tennis Court Road
Old Addenbrookes Site
Cambridge, CB2 1GA, UK
Tel: +44 (1223) 333639 (including answerphone)
FAX: +44 (1223) 766002
e-mail: lcp2@mole.bio.cam.ac.uk
Visit my WWW page at http://www.bio.cam.ac.uk/proj/adr/PNAC/pnac.html