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Protein Sequencing

  • Bryan J. Smith
  • John R. Chapman
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

It is approaching half a century since Edman developed a chemistry for the determination of the N-terminus of a protein or peptide (1), and thts chemistry is still being used today. in the intervening decades various ancillary techniques have come (and some have gone) that have allowed the field to develop. Apart from the automation of Edman chemistry itself, the most sigmticant advances have been improvements in sample preparation and the advent of mass spectrometric techniques. in the early days the task of sequencing a protem was a signnificant one, limited to those protems that could be prepared in sufficiently large amounts. Then, the aim (other than to develop the technology) was to obtam the full sequence of the protein in question to begin to understand how protems were structured, which was done by use of Edman chemistry exclusively. Currently, the situation is different, and it is usually necessary to obtam only partial protein sequence from the protein itself From the partial sequence, suitable ohgonucleotides can be designed and used to clone the corresponding gene, which can be rapidly analyzed to yield the full sequence, For identification of a protein the partial sequence may need to be only 3–5 residues long Such short sequences, known as “sequence tags,” are currently of importance in the study of the “proteome,” the set of protems that is expressed by the genome. Studies of the proteome follow the results of genome sequencing, which are giving an explosion of information. The sequence of the genome suggests what proteins might be made, but does not prove which are actually expressed in any gtven tissue(s), how they are regulated, or what their functions e For these purposes there is a need to separate and identify mdividual proteins, then to correlate then presence and modification with function.

Keywords

Sodium Dodecyl Sulfate Protein Spot Cyanogen Bromide MALDI Mass Spectrometry Mass Spectrometric Technique 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Edman, P (1950) Method for determmatlon of the ammo acid sequence in peptides Actu Chem Scand 4, 283–293CrossRefGoogle Scholar
  2. 2.
    Smith, B J and Tempst, P (1996) Strategies for handling polypeptldes on a microscale, in Methods in Molecular Biology, vol 64 Protein Sequencing Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 1–16CrossRefGoogle Scholar
  3. 3.
    Chang, H W and Bock, E (1980) Pitfalls in the use of commercial nomomc detergents for the solublllsatlon of integral membrane proteins sulfhydryl oxldlsing contaminants and their ehmmatlon Anal Bzochem 104, 112–117CrossRefGoogle Scholar
  4. 4.
    Ogorzalek Loo, R R, Dales, N, and Andrews P C (1996) The effect of detergents on proteins analyzed by electrospray ionization, in Methods in Molecular Bzology, vol 61 Protem and Peptzde Analyszs by Mass Spectrometry (Chapman, J R, ed), Humana, Totowa, NJ, pp 141–160Google Scholar
  5. 5.
    Bornsen, K O, Gass, M A, Bruin, G J, von Adnchem, J H A, Biro, M C, Kresbach, G M, and Ehrat, M (1997) Influence of solvents and detergents on matrix-assisted laser desorptlon/lomzatlon mass spectrometry measurements of proteins and ollgonucleotldes Rapid Commun Mass Spectrom 11,603–609PubMedCrossRefGoogle Scholar
  6. 6.
    Schey, K L (1996) Hydrophobic proteins and peptldes analyzed by matrix-assisted laser desorptlonllomzatlon, in Methods WI Molecular Bzology, vol 61 Protein and Peptzde,4nalysls by Muss Spectrometry (Chapman, J R, ed), Humana, Totowa, NJ, pp 227–230CrossRefGoogle Scholar
  7. 7.
    Phllp, R J (1996) Preparation of peptides for mlcrosequencing from proteins in polyacrylamlde gels, in The Proteain Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 393–398CrossRefGoogle Scholar
  8. 8.
    Ward, M (1996) In situ chemical and enzymatic digestions of proteins nnmoblhzed on miniature hydrophobic columns, in The Protezn Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 399–403CrossRefGoogle Scholar
  9. 9.
    Femandez, J and Mlsche, S M (1996) Enzymatic digestion of membrane-bound proteins for peptlde mapping and internal sequence analysis, in The Protein Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 405–414CrossRefGoogle Scholar
  10. 10.
    Stone, K L and Wllhams, K R (1996) Enzymatic digestion of proteins in solution and in SDS polyacrylamlde gels, in The Protein Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 415–425CrossRefGoogle Scholar
  11. 11.
    Judd, R C (1996) SDS-polyacrylamide gel electrophoresls of peptides, in The Proteain Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 101–107CrossRefGoogle Scholar
  12. 12.
    Judd, R C (1996) Peptide mapping by sodium dodecylsulfate-polyacrylamlde gel electrophoresis, in The Protein Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ,pp 447–451CrossRefGoogle Scholar
  13. 13.
    Judd, R C (1996) Peptide mapping by high-performance llquld chromatography, in The Proteain Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 453–455CrossRefGoogle Scholar
  14. 14.
    Shaw, C (1996) Reverse-phase HPLC purlficatlon of peptldes from natural sources for structural analysis, in Methods in Molecular Bzologv, vol 64 Protein Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 101–107CrossRefGoogle Scholar
  15. 15.
    Smith, A J (1996) Analytlcal and micropreparatlve capillary electrophoresls of peptides, in Methods in Molecular Bzology, vol 64 Protem Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 91–99CrossRefGoogle Scholar
  16. 16.
    Sutton, C W and O’Nelll, J A (1996) Preparation of glycopeptldes, in Methods in Molecular Biologv, vol 64 Protein Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 73–79CrossRefGoogle Scholar
  17. 17.
    Hayashl, T H and Sasagawa, T (1996) Selective lsolatlon of the carboxy terminal peptide from a peptlde, in Methods in Molecular Biology, vol 64 Protem Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 81–83Google Scholar
  18. 18.
    Hayashl, T H and Sasagawa, T (1996) Selective lsolatlon oftheammo terminal peptide from an a-ammo blocked protein, in Methods in Molecular Biology, vol 64 Protein Sequencing Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 85–89Google Scholar
  19. 19.
    Tawfik, D (1996) Side-chain selective chemical modlficatlons of proteins, in The Protezn Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 349–351CrossRefGoogle Scholar
  20. 20.
    Spelcher, D W (1996) Chemical modification of proteins, in Current Protocols in Protezn Sczence, vol 1 (Cohgan, J E, Dunn, B M, Ploegh, H H, Spelcher, D W, and Wingfield, P T, eds), Wiley, New York, pp 15 0 1,15 0 2Google Scholar
  21. 21.
    Came, A F and U, S (1996) Chemical modlficatlon of proteins for sequence analysis, in Methods in Molecular Biology, vol 64 Protein Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 271–284Google Scholar
  22. 22.
    Glazer, A N, Delange, R J, and Slgman, D S (1975) Chemical Modzficatzon of Proteins Selected Methods and Analytzcal Procedures North Holland Pub Co Amsterdam, Oxford, American Elsevler Pub Co Inc, NYGoogle Scholar
  23. 23.
    Crankshaw, M W and Grant, G A (1996) Modlficatlon of cysteme, in Current Protocols in Protein Sczence, vol 1 (Cohgan, J E, Dunn, B M, Ploegh, H H, Spelcher, D W, and Wingfield, P T, eds) Wiley, New York, pp 1511–15118Google Scholar
  24. 24.
    Hounsell, E F, Davies, M J and Smith, K D (1996) Enzymatic release of O-and N-linked ohgosaccharlde chains, in The Protein Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 657,658CrossRefGoogle Scholar
  25. 25.
    Hounsell, E F, Davies, M J, and Smith, K D (1996) Chemical release of O-linked ohgosaccharlde chains, in The Protezn Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 647,648CrossRefGoogle Scholar
  26. 26.
    Mlzuochl, T and Hounsell, E F (1996) Release of N-linked ohgosaccharlde chains by hydrazmolysls, in The Protein Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 653–656Google Scholar
  27. 27.
    Colyer, J (1996) Analysing protein phosphorylatlon, in The Protem Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 501–506CrossRefGoogle Scholar
  28. 28.
    Hlrano, H, Komatsu, S, and Tsunasawa, S (1996) On-membrane deblocking of protems, in Methods in Molecular Blologv, vol 64 Protem Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 285–292Google Scholar
  29. 29.
    Walker, J M and Sweeney, P J (1996) Removal of pyroglutamlc acid residues from the N-terminus of peptldes and proteins, in The Protem Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 525–527CrossRefGoogle Scholar
  30. 30.
    Fenn, J B, Mann, M, Meng, C K, Wong, S F, and Whltehouse, C M (1990) Electrospray lomzatlon—priciples and practice Mass Spectrom Rev 9, 37–70CrossRefGoogle Scholar
  31. 31.
    Hillenkamp, F, Karas, M, Beavls, R C, and Chalt, B T (1991) Matrix-assisted laser desorption/iomzatlon of biopolymers Anal Chem 63, 1193A–1203APubMedCrossRefGoogle Scholar
  32. 32.
    Bennett, K L, Hick, L A, Truscott, R J W, Shlel, M M, and Smith, S V (1995) Optimum condltlons for electrospray mass spectrometry of a monoclonal antibody J Mass Spectrom 30,769–771CrossRefGoogle Scholar
  33. 33.
    He, S, Pan, S, Wu, K, Amster, I J, and Orlando, R (1995) Analysis of normal fetal eye lens crystallms by high-performance hquld chromatography/mass spectrometry J Mass Spectrom 30,424–431CrossRefGoogle Scholar
  34. 34.
    Light-Wahl, K J, Schwartz, B L, and Smith, R D (1994) Observation of the noncovalent quaternary assoclatlons of proteins by electrospray lomzatlon mass spectrometry J Am Chem Sot 116,5271–5278CrossRefGoogle Scholar
  35. 35.
    Jespersen, S, Koedam, J A, Hoogerbrugge, C M, TJaden, U R, van der Greef, J, and Van den Brande, J L (1996) Characterrzatlon of O-glycosylated precursors of msulmlike growth factor II by matrix-assisted laser desorptlon/lomzatlon mass spectrometry J Mass Spectrom 31, 893–900PubMedCrossRefGoogle Scholar
  36. 36.
    Wada, Y (1996) Structural analysis of protein vanants, in Methods in Molecular Bzology, vol 61 Protew and Peptrde Analysis by Mass Spectrometry (Chapman, J R, ed), Humana, Totowa, NJ, pp 101–113CrossRefGoogle Scholar
  37. 37.
    Cottrell, J S and Sutton, C W (1996) The ldentlficatlon ofelectrophoretlcally separated proteins by peptlde mass fingerprmting, in Methods in Molecular Biology, vol 61 Protem and Peptzde Analysis by Mass Spectrometry (Chapman, J R, ed), Humana, Totowa, NJ, pp 67–82CrossRefGoogle Scholar
  38. 38.
    Aebersold, R H, Leavltt, J, Saavedra, R A, Hood, L E, and Kent, S B H (1987) Internal ammo acid sequence analysis of proteins separated by one-or two-dlmenslonal gel electrophoresls after in situ protease digestion on nitrocellulose Proc Natl Acad Sci USA 84, 6970–6974PubMedCrossRefGoogle Scholar
  39. 39.
    Henderson, L E, Oroszlan, S, and Koingsberg, W (1979) A mlcromethod for complete removal of dodecyl sulfate from proteins by ion-pair extractlon Anal Bzochem 93,153–157CrossRefGoogle Scholar
  40. 40.
    Coull, J M and Pappm, D J C (1990) A rapid fluorescent staining procedure for proteins electroblotted onto PVDF membranes J Protein Chem 9,259,260Google Scholar
  41. 41.
    Ortiz, M L, Calero, M, Patron, C F, Castellanos, L, and Mendez, E (1992) Imidazole-SDS-zinc reverse staming of protems in gels containing or not SDS and microsequence of mdlvldual unmodtfied electroblotted proteins FEBS Lett 296,300–304PubMedCrossRefGoogle Scholar
  42. 42.
    Vorm, O, Chatt, B T, and Roepstorff, P (1993) Mass spectrometry of protein samples contaming detergents Proceedzngs of the 41 stAnnual Conference on Mass Spectrometry and Allzed Topzcs, San Francisco, CA, pp. 621Google Scholar
  43. 43.
    Nelson, R W, Dogruel, D, Krone, J R, and Wilhams, P (1995) Pepttde charactertzation using btoreacttve mass spectrometer probe tips Rapzd Commun MassSpectrom 9,1380–1385CrossRefGoogle Scholar
  44. 44.
    Covey, T (1996) Liquid chromatography/mass spectrometry for the analysis of protein digests, in Methods in Molecular Biology, vol 61 Protein and Peptzde Analyszs by Mass Spectrometry (Chapman, J R, ed), Humana, Totowa, NJ, pp, 83–100CrossRefGoogle Scholar
  45. 45.
    Mann, M and Wtlm, M (1994) Error-tolerant tdenttficatron of peptides in sequence databases by peptide sequence tags Anal Chem 66,4390–4399PubMedCrossRefGoogle Scholar
  46. 46.
    Bonner, R and Shushan, B (1995) The charactertzatlon of protems and pepttdes by automated methods Rapzd Commun MassSpectrom 9,1067–1076CrossRefGoogle Scholar
  47. 47.
    Bonner, R and Shushan, B (1995) Error-tolerant protem database searching using peptide product-ton spectra Rapzd Commun MassSpectrom 9, 1077–1080CrossRefGoogle Scholar
  48. 48.
    Barber, M, Bordoh, R S, Ellitt, G J, Sedgwtck, R D, and Tyler, A N (1982) Fastatom bombardment mass spectrometry Anal Chem 54,645A–657ACrossRefGoogle Scholar
  49. 49.
    Carr, S A, Huddleston, M J, and Bean, M (1993) Selective tdenttficatton and differentiation of N- and O-linked ohgosaccharides in glycoprotems by hqmd chromatographymas spectrometry Protezn Scz 2, 183–196CrossRefGoogle Scholar
  50. 50.
    Huddleston, M J, Bean, M, and Carr, S. A (1993) Collisional fragmentation of glycopeptides by electrospray tomzatlon LC/MS and LC/MS/MS methods for selecttve detection of glycopeptides in protem digests Anal Chem 65,877–882PubMedCrossRefGoogle Scholar
  51. 51.
    Huddleston, M J., Annan, R S, Bean, M, and Carr, S A (1993) Selective detectton of the thr-, ser-, and tyr-phosphopepttdes in complex digests by electrospray LC-MS J Am Sot MassSpectrom 4,710–717CrossRefGoogle Scholar
  52. 52.
    Ding, J, Burkhary, W, and Kassel, D B (1994) Identtficatton of phosphorylated pepttdes from complex mixtures using neganve-ton orifice-potential stepping and captllary liquid chromatographylelectrospray tomzatton mass spectrometry. Rapzd Commun Mass Spectrom 8,94–98CrossRefGoogle Scholar
  53. 53.
    Covey, T R, Shushan, B, Bonner, R, Schroder, W, and Hucho, F (1991) LUMS and LC/MS/MS screening for the sites of post-translational modtfication in proteins, in Methods in Protezn Sequence Analyszs (Jornvall, H, Hoog, J-O, and Gustavsson, A-M, eds), Blrkhauser Verlag, Basel, Switzerland, pp 249–256Google Scholar
  54. 54.
    Feng, F, Bell, A, Dumas, F., and Konishi, Y (1990) Reductton/alkylatton plus mass spectrometry a fast and simple method for accurate counting of cystemes, dtsultide bridges and free SH groups in proteins Proceedings of the 38 th Annual Conference on Mass Spectrometry and Allied Topics, Tuczon, AZ, pp 273,274Google Scholar
  55. 55.
    Sun, Y., Bauer, M D, Keough, T W, and Lacey, M. P (1996) Dtsulfide bond location in proteins, in Methods in Molecular Bzology, vol 61 Protezn and Peptzde Analyszs by Mass Spectrometry (Chapman, J R, ed), Humana, Totowa, NJ, pp 185–210CrossRefGoogle Scholar
  56. 56.
    Sun, Y, Zhou, Z, and Smith, D L (1989) Locatton of dtsulfide bonds in proteins by parttal acid hydrolysis and mass spectrometry, in Technzques in Proteain Chemzstry (Hugh T E, ed), Academtc, New York, pp. 176–185Google Scholar
  57. 57.
    Yazdanparast, R, Andrews, P C, Smith, D L, and Dtxon, J E (1987) Assignment of drsulfide bonds in proteins by fast atom bombardment mass spectrometry J Bzol Chem 262,2507–2513Google Scholar
  58. 58.
    Sun, Y and Smith, D L (1988) Identification of dlsulflde-contaming peptldes by performic acid oxldatlon and mass spectrometry Anal Bzochem 197, 69–76CrossRefGoogle Scholar
  59. 59.
    Alken, A and Learmath, M (1996) Analysis of sites of protein phosphorylatlon, in Methods in Molecular Bzology, vol 64 Protem Sequencing Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 293–306Google Scholar
  60. 60.
    Chalt, B T, Wang, R, Beavls, R C, and Kent, S B H (1993) Protein ladder sequencing Science 262, 89–92CrossRefGoogle Scholar
  61. 61.
    Bartlet-Jones, M, Jeffery, W A, Hansen, H F, and Pappm, D J C (1994) Peptlde ladder sequencing by mass spectrometry using a novel, volatile degradation reagent Rapzd Commun Mass Spectrom 8, 737–742CrossRefGoogle Scholar
  62. 62.
    Inghs, A (1991) Chemical procedures for C-terminal sequencing of peptldes and proteins Analyt Bzochem 195, 183–196CrossRefGoogle Scholar
  63. 63.
    Casagranda, F and Wllshlre, J F K (1996) Enzymatic and chemical methods for manual C-terminal peptlde sequencing In Methods rn Molecular Biology, vol 64 Protein Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 243–257Google Scholar
  64. 64.
    Bailey, J M and Iller, C G (1996) Automated methods for C-terminal protein sequencing In Methods in Molecular Btology, vol 64 Protem Sequencing Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 259–269CrossRefGoogle Scholar
  65. 65.
    Ambler, R P (1972) Enzymatic hydrolysis with carboxypeptldases, in Methods in EnzymoZ, vol 25 (Hers, C H W and Tlmasheff, S N, eds), Academic, New York, pp 143–154Google Scholar
  66. 66.
    Walker, J M and Wmder, J S (1996) C-terminal sequence analysis with carboxypeptldase Y, in The Protezn Protocols Handbook (Walker, J M, ed), Humana, Totowa, NJ, pp 569–571CrossRefGoogle Scholar
  67. 67.
    Busch, K L, Ghsh, G L, and McLuckey, S A (1988) Muss SpectrometryiMass Spectrometry Technzques and Applrcatlons of Tandem Mass Spectrometry VCH, Wemhelm, GermanyGoogle Scholar
  68. 68.
    Spengler, B (1996) New Instrumental approaches to colllslon-induced dlssoclatlon using a time-of-flight instrument, in Methods in Molecular Biology, vol 61 Proteain and Pepttde Analyszs by Mass Spectrometry (Chapman, J R, ed), Humana, Totowa, NJ,pp 43–56CrossRefGoogle Scholar
  69. 69.
    Johnson, R S, Martin, S A, and Blemann, K (1988) Colhslon-induced fragmentation of (M + H)+ ions of peptldes Side chain specific sequence ions Int J Mass Spectrom Ion Processes 86, 137–154CrossRefGoogle Scholar
  70. 70.
    Johnson, R S, Martin, S A, Blemann, K, Stults, J T, and Watson, J T (1987) Novel fragmentation processes of peptldes by colhslon-induced decomposltlon in a tandem mass spectrometer differentiation of leucme and lsoleucme Anal Chem 59,2621–2625PubMedCrossRefGoogle Scholar
  71. 71.
    Blemann, K (1990) Sequencing of peptldes by tandem mass spectrometry and hlghenergy colllslon-Induced dlssoclatlon Methods Enzymol 193,455–479CrossRefGoogle Scholar
  72. 72.
    Blemann, K (1988) Contrlbutlons of mass spectrometry to peptlde and protein structure Biamed Envwon Mass Spectrom 16,99–111CrossRefGoogle Scholar
  73. 73.
    Blemann, K and Scoble, H A (1987) Characterlzatlon by tandem mass spectrometry Science 237, 992–998CrossRefGoogle Scholar
  74. 74.
    Gibson, B W and Cohen, P (1990) Liquid secondary Ion mass spectrometry ofphosphorylated and sulfated peptldes and proteins Methods Enzymol 193,480–501PubMedCrossRefGoogle Scholar
  75. 75.
    Zala, J and Blemann, K (1995) Comparison of charged derivatives for high energy collision-induced dlssoclatlon mass spectrometry J Am Sot Mass Spectrom 6, 428–436CrossRefGoogle Scholar
  76. 76.
    Covey, T R, Huang, E C, and Hemon, J D (1991) Structural characterlzatlon of protein tryptlc peptldes via hquld chromatography/mass spectrometry and collision-Induced dlssoclatlon of their doubly charged molecular ions Anal Chem 63,1193–1200PubMedCrossRefGoogle Scholar
  77. 77.
    DavIdson, I (1996) Hydrolysis of samples for ammo acid analysis, in Methods in Molecular Biology, vol 64 Proteain Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 119–129CrossRefGoogle Scholar
  78. 78.
    Irvine, G B (1996) Ammo acid analysis precolumn derlvatlzatlon methods, in Methods in Molecular Bzology, vol 64 Protem Sequencing Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 131–138CrossRefGoogle Scholar
  79. 79.
    Smith, A J (1996) Postcolumn ammo acid analysis, in Methods in Molecular Bzology, vol 64 Proteain Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 139–146CrossRefGoogle Scholar
  80. 80.
    Nakagawa, S and Fukuda, T (1989) Direct ammo acid analysis of proteins electroblotted onto polyvmyhdene difluorlde membrane from sodium dodecylsulfate-polyacrylamide gel Analyt Blochem 181, 75–78CrossRefGoogle Scholar
  81. 81.
    Hobohm, U, Houthaeve, T, and Sander, C (1994) Ammo acid analysis and protein database composltlon search as a fast and mexpenslve method to identify proteins Anal Blochem 222,202–209CrossRefGoogle Scholar
  82. 82.
    Turck, C W (1996) High performance hquld chromatography on-lme derlvatlve spectroscopy for the characterlsatlon of peptides with aromatic ammo acid residues, in Methods it Molecular Bzology, vol 64 Protem Sequenczng Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 109–117Google Scholar
  83. 83.
    Bray, M R, Camere, A D, and Clarke, A J (1994) Quantltatlon of tryptophan and tyrosme residues in proteins by fourth derlvatlve spectroscopy Analyt Blochem 221, 278–284CrossRefGoogle Scholar
  84. 84.
    Mach, H and Mlddaugh, C R (1994) Srmultaneous momtoring of the environment of tryptophan, tyrosme and phenylalamne residues in proteins by near-ultravlolet second derivative spectroscopy Anal Blochem 222,323–331CrossRefGoogle Scholar
  85. 85.
    Smith, B J and Wheeler, C (1996) Enzymatic cleavage of polypeptldes, in Methods in Molecular Bzologv, vol 64 Protew Sequencing Protocols (Smith, B J, ed), Humana, Totowa, NJ, pp 43–55CrossRefGoogle Scholar
  86. 86.
    Bornstem, P and Balian, G (1977) Cleavage at Asn-Gly bonds with hydroxylamme Methods Enzymol 47, 143–145Google Scholar
  87. 87.
    Inghs, A S (1983) Cleavage at aspartlc acid Methods Enzymol 91,324–332CrossRefGoogle Scholar
  88. 88.
    Swenson, C A and Fredrickson, R S (1992) Interaction of troponin C and troponin C fragments with troponin I and the troponm I inhibitory peptlde Biochemzstry 31, 3420–3427CrossRefGoogle Scholar
  89. 89.
    Fontana, A and Gross, E (1986) Fragmentation of polypeptldes by chemical methods, in Practwal Protein Chemistry A Handbook (Darbre, A, ed), Wiley, Chichester, pp 67–120Google Scholar
  90. 90.
    Huang, H V, Bond, M W, Hunkaplllar, M W, and Hood, L E (1983) Cleavage at tryptophanyl residues with dlmethyl sulfoxlde-hydrochloric acid and cyanogen bromide Methods Enzymol 91,318–324PubMedCrossRefGoogle Scholar
  91. 91.
    Kanda, F, Yoshlda, S, Okumura, T, and Takamatsu, T (1995) Asparagmyl endopeptldase mapping of protems with subsequent matrix-assisted laser desorptlon/lomzatlon mass spectrometry Raped Commun Mass Spectrom 9,1095–1100CrossRefGoogle Scholar

Copyright information

© Humana Press Inc , Totowa, NJ. 1998

Authors and Affiliations

  • Bryan J. Smith
    • 1
  • John R. Chapman
    • 2
  1. 1.Celltech TherapeuttcsSloughUK
  2. 2.CheshireUK

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