31:
323:, the reagent for the Edman degradation, can also be used. The same questions apply here as in the determination of amino acid composition, with the exception that no stain is needed, as the reagents produce coloured derivatives and only qualitative analysis is required. So the amino acid does not have to be eluted from the chromatography column, just compared with a standard. Another consideration to take into account is that, since any amine groups will have reacted with the labelling reagent, ion exchange chromatography cannot be used, and
246:
96:
496:
570:, which cleaves selectively on the C-terminal side of Lysine or Arginine residues, is the most commonly used protease. Its advantages include i) the frequency of Lys and Arg residues in proteins, ii) the high specificity of the enzyme, iii) the stability of the enzyme and iv) the suitability of tryptic peptides for mass spectrometry.
678:
The protein’s whole mass is the sum of the masses of its amino-acid residues plus the mass of a water molecule and adjusted for any post-translational modifications. Although proteins ionize less well than the peptides derived from them, a protein in solution may be able to be subjected to ESI-MS and
603:
Information including the measured mass of the putative peptide ions and those of their fragment ions is then matched against calculated mass values from the conceptual (in-silico) proteolysis and fragmentation of databases of protein sequences. A successful match will be found if its score exceeds a
712:
of genes and may be further modified. These processes are sufficiently understood to use computer algorithms to automate predictions of protein sequences from DNA sequences, such as from whole-genome DNA-sequencing projects, and have led to the generation of large databases of protein sequences such
366:
is a very important reaction for protein sequencing, because it allows the ordered amino acid composition of a protein to be discovered. Automated Edman sequencers are now in widespread use, and are able to sequence peptides up to approximately 50 amino acids long. A reaction scheme for sequencing a
347:
to a solution of the protein, take samples at regular intervals, and determine the terminal amino acid by analysing a plot of amino acid concentrations against time. This method will be very useful in the case of polypeptides and protein-blocked N termini. C-terminal sequencing would greatly help in
687:
Proteolysis does not always yield a set of readily analyzable peptides covering the entire sequence of POI. The fragmentation of peptides in the mass spectrometer often does not yield ions corresponding to cleavage at each peptide bond. Thus, the deduced sequence for each peptide is not necessarily
539:
Protein identification is the process of assigning a name to a protein of interest (POI), based on its amino-acid sequence. Typically, only part of the protein’s sequence needs to be determined experimentally in order to identify the protein with reference to databases of protein sequences deduced
225:
Pre-column derivatization may use the Edman reagent to produce a derivative that is detected by UV light. Greater sensitivity is achieved using a reagent that generates a fluorescent derivative. The derivatized amino acids are subjected to reversed phase chromatography, typically using a C8 or C18
213:
can be used for this; it gives a yellow colour when reacted with proline, and a vivid purple with other amino acids. The concentration of amino acid is proportional to the absorbance of the resulting solution. With very small quantities, down to 10 pmol, fluorescent derivatives can be formed using
103:
It is often desirable to know the unordered amino acid composition of a protein prior to attempting to find the ordered sequence, as this knowledge can be used to facilitate the discovery of errors in the sequencing process or to distinguish between ambiguous results. Knowledge of the frequency of
306:
There are many different reagents which can be used to label terminal amino acids. They all react with amine groups and will therefore also bind to amine groups in the side chains of amino acids such as lysine - for this reason it is necessary to be careful in interpreting chromatograms to ensure
644:
The peptides matched during protein identification do not necessarily include the N- or C-termini predicted for the matched protein. This may result from the N- or C-terminal peptides being difficult to identify by MS (e.g. being either too short or too long), being post-translationally modified
691:
Because the Edman degradation proceeds from the N-terminus of the protein, it will not work if the N-terminus has been chemically modified (e.g. by acetylation or formation of
Pyroglutamic acid). Edman degradation is generally not useful to determine the positions of disulfide bridges. It also
615:
A diagram of the matched peptides on the sequence of the identified protein is often used to show the sequence coverage (% of the protein detected as peptides). Where the POI is thought to be significantly smaller than the matched protein, the diagram may suggest whether the POI is an N- or
512:
is a machine that performs Edman degradation in an automated manner. A sample of the protein or peptide is immobilized in the reaction vessel of the protein sequenator and the Edman degradation is performed. Each cycle releases and derivatises one amino acid from the protein or peptide's
657:
Whilst detailed comparison of the MS data with predictions based on the known protein sequence may be used to define post-translational modifications, targeted approaches to data acquisition may also be used. For instance, specific enrichment of phosphopeptides may assist in identifying
163:) are degraded. To circumvent this problem, Biochemistry Online suggests heating separate samples for different times, analysing each resulting solution, and extrapolating back to zero hydrolysis time. Rastall suggests a variety of reagents to prevent or reduce degradation, such as
679:
its mass measured to an accuracy of 1 part in 20,000 or better. This is often sufficient to confirm the termini (thus that the protein’s measured mass matches that predicted from its sequence) and infer the presence or absence of many post-translational modifications.
424:
Peptides longer than about 50–70 amino acids long cannot be sequenced reliably by the Edman degradation. Because of this, long protein chains need to be broken up into small fragments that can then be sequenced individually. Digestion is done either by
540:
from the DNA sequences of their genes. Further protein characterization may include confirmation of the actual N- and C-termini of the POI, determination of sequence variants and identification of any post-translational modifications present.
759:
as a basis for creating k-time programs, programs that run exactly k times before self-destructing. Such a thing is impossible to build purely in software because all software is inherently clonable an unlimited number of times.
611:
Software packages usually generate a report showing the identity (accession code) of each identified protein, its matching score, and provide a measure of the relative strength of the matching where multiple proteins are
604:
threshold based on the analysis parameters. Even if the actual protein is not represented in the database, error-tolerant matching allows for the putative identification of a protein based on similarity to
209:. Once the amino acids have been separated, their respective quantities are determined by adding a reagent that will form a coloured derivative. If the amounts of amino acids are in excess of 10 nmol,
234:
gradient. The eluting amino acids are detected using a UV or fluorescence detector and the peak areas compared with those for derivatised standards in order to quantify each amino acid in the sample.
108:
to use for digestion of the protein. The misincorporation of low levels of non-standard amino acids (e.g. norleucine) into proteins may also be determined. A generalized method often referred to as
57:. Typically, partial sequencing of a protein provides sufficient information (one or more sequence tags) to identify it with reference to databases of protein sequences derived from the conceptual
581:) but further fragmentation of the peptides inside the mass spectrometer is often used to gain information about the peptides’ sequences. Alternatively, peptides may be desalted and separated by
492:, which can be washed off and identified by chromatography, and the cycle can be repeated. The efficiency of each step is about 98%, which allows about 50 amino acids to be reliably determined.
201:
An example of the ion-exchange chromatography is given by the NTRC using sulfonated polystyrene as a matrix, adding the amino acids in acid solution and passing a buffer of steadily increasing
80:(sequencer). Mass spectrometry methods are now the most widely used for protein sequencing and identification but Edman degradation remains a valuable tool for characterizing a protein's
645:(e.g. N-terminal acetylation) or genuinely differing from the prediction. Post-translational modifications or truncated termini may be identified by closer examination of the data (i.e.
720:
Historically, short protein sequences (10 to 15 residues) determined by Edman degradation were back-translated into DNA sequences that could be used as probes or primers to isolate
1191:
348:
verifying the primary structures of proteins predicted from DNA sequences and to detect any posttranslational processing of gene products from known codon sequences.
724:
of the corresponding gene or complementary DNA. The sequence of the cloned DNA was then determined and used to deduce the full amino-acid sequence of the protein.
574:
1244:
1083:
1331:
280:
chain is useful for two reasons: to aid the ordering of individual peptide fragments' sequences into a whole chain, and because the first round of
922:
Shevchenko A, Tomas H, Havlis J, Olsen JV, Mann M (2006). "In-gel digestion for mass spectrometric characterization of proteins and proteomes".
1326:
582:
195:
30:
865:
517:-terminus and the released amino-acid derivative is then identified by HPLC. The sequencing process is done repetitively for the whole
986:
441:. Different enzymes give different cleavage patterns, and the overlap between fragments can be used to construct an overall sequence.
708:
of messenger RNA (mRNA) with the protein sequence deriving from the sequence of codons in the mRNA. The mRNA is itself formed by the
688:
complete. The standard methods of fragmentation do not distinguish between leucine and isoleucine residues since they are isomeric.
577:
mass spectrometry. Direct measurement of the masses of the peptides may provide sufficient information to identify the protein (see
174:
to protect tryptophan and tyrosine from attack by chlorine, and pre-oxidising cysteine. He also suggests measuring the quantity of
343:
amino acid analysis is much smaller than the number of available methods of N-terminal analysis. The most common method is to add
1181:
1161:
1142:
633:
54:
663:
593:
563:
The isolated POI may be chemically modified to stabilise
Cysteine residues (e.g. S-amidomethylation or S-carboxymethylation).
1357:
1171:
1076:
667:
1219:
592:
Depending on the type of mass spectrometer, fragmentation of peptide ions may occur via a variety of mechanisms such as
1318:
1224:
1152:
589:
source. LC-ESI-MS may provide more information than MALDI-MS for protein identification but uses more instrument time.
578:
312:
258:
191:
736:
626:
534:
1176:
1166:
1156:
1118:
530:
194:
then derivatized to facilitate their detection. More commonly, the amino acids are derivatized then resolved by
1196:
1069:
454:
636:
or chemical modifications. It may provide additional evidence for protein identifications performed as above.
709:
586:
139:
groups may require longer heating periods. However, these conditions are so vigorous that some amino acids (
1267:
1229:
717:. Predicted protein sequences are an important resource for protein identification by mass spectrometry.
489:
1352:
1186:
967:"Preparation of proteins and peptides for mass spectrometry analysis in a bottom-up proteomics workflow"
744:
705:
227:
58:
692:
requires peptide amounts of 1 picomole or above for discernible results, making it less sensitive than
600:(PSD). In each case, the pattern of fragment ions of a peptide provides information about its sequence.
95:
245:
1234:
1113:
466:
320:
1201:
1147:
1138:
662:
sites in a protein. Alternative methods of peptide fragmentation in the mass spectrometer, such as
42:
1049:
947:
284:
is often contaminated by impurities and therefore does not give an accurate determination of the
1288:
1041:
1002:
982:
939:
904:
861:
836:
740:
721:
605:
521:
until the entire measurable sequence is established or for a pre-determined number of cycles.
380:
363:
357:
281:
206:
132:
625:
The pattern of fragmentation of a peptide allows for direct determination of its sequence by
394:
Separate and purify the individual chains of the protein complex, if there are more than one.
1303:
1033:
992:
974:
931:
894:
826:
462:
438:
384:
372:
367:
protein by the Edman degradation follows; some of the steps are elaborated on subsequently.
344:
179:
649:
sequencing). A repeat digest using a protease of different specificity may also be useful.
1272:
1096:
659:
470:
388:
316:
1211:
1128:
997:
966:
899:
882:
784:
774:
732:
557:
474:
376:
328:
324:
965:
Gundry RL, White MY, Murray CI, Kane LA, Fu Q, Stanley BA, Van Eyk JE (October 2009).
831:
814:
813:
Bogosian G, Violand BN, Dorward-King EJ, Workman WE, Jung PE, Kane JF (January 1989).
632:. This sequence may be used to match databases of protein sequences or to investigate
1346:
1308:
1298:
1239:
794:
789:
779:
485:
426:
296:
React the peptide with a reagent that will selectively label the terminal amino acid.
219:
1293:
1262:
1123:
1053:
951:
215:
1061:
1024:
Steen H, Mann M (September 2004). "The ABC's (and XYZ's) of peptide sequencing".
978:
855:
99:
Protein sequence interpretation: a scheme new protein to be engineered in a yeast
17:
608:
proteins. A variety of software packages are available to perform this analysis.
518:
495:
205:
through the column. Amino acids are eluted when the pH reaches their respective
136:
815:"Biosynthesis and incorporation into protein of norleucine by Escherichia coli"
573:
The peptides may be desalted to remove ionizable contaminants and subjected to
1105:
769:
597:
450:
340:
270:
152:
128:
81:
481:
480:
The terminal amino acid can then be selectively detached by the addition of
458:
210:
156:
144:
1045:
1006:
943:
935:
908:
840:
553:
302:
Determine the amino acid by chromatography and comparison with standards.
160:
148:
105:
1092:
756:
714:
567:
469:(PITC), is added to the adsorbed peptide, together with a mildly basic
430:
277:
175:
167:
116:
Hydrolyse a known quantity of protein into its constituent amino acids.
50:
46:
434:
231:
171:
140:
1037:
135:
to 100–110 °C for 24 hours or longer. Proteins with many bulky
566:
The POI is digested with a specific protease to generate peptides.
307:
that the right spot is chosen. Two of the more common reagents are
1254:
494:
164:
94:
29:
477:. This reacts with the amine group of the N-terminal amino acid.
62:
1065:
743:), or search databases using peptide or protein sequences (see
735:
tools exist to assist with interpretation of mass spectra (see
53:. This may serve to identify the protein or characterize its
499:
A Beckman-Coulter Porton LF3000G protein sequencing machine
202:
548:
A general scheme for protein identification is described.
403:
Break each chain into fragments under 50 amino acids long.
693:
69:
854:
Michail A. Alterman; Peter
Hunziker (2 December 2011).
68:
The two major direct methods of protein sequencing are
391:
may be necessary to prevent the bonds from re-forming.
34:
Using a
Beckman-Spinco Protein-Peptide Sequencer, 1970
104:
certain amino acids may also be used to choose which
112:
for determining amino acid frequency is as follows:
1317:
1281:
1253:
1210:
1104:
397:
Determine the amino acid composition of each chain.
755:The difficulty of protein sequencing was recently
265:total acid hydrolysis of the dinitrophenyl peptide
131:is done by heating a sample of the protein in 6 M
119:Separate and quantify the amino acids in some way.
400:Determine the terminal amino acids of each chain.
27:Sequencing of amino acid arrangement in a protein
739:), to compare or analyze protein sequences (see
670:, may give complementary sequence information.
585:and introduced into a mass spectrometer via an
288:-terminal amino acid. A generalised method for
249:Sanger's method of peptide end-group analysis:
616:C-terminal fragment of the identified protein.
415:Construct the sequence of the overall protein.
1077:
8:
412:Repeat with a different pattern of cleavage.
41:is the practical process of determining the
1084:
1070:
1062:
857:Amino Acid Analysis: Methods and Protocols
996:
898:
830:
409:Determine the sequence of each fragment.
244:
805:
292:-terminal amino acid analysis follows:
269:Determining which amino acid forms the
77:
1327:Photoactivated localization microscopy
1245:Protein–protein interaction prediction
971:Current Protocols in Molecular Biology
73:
1026:Nature Reviews Molecular Cell Biology
704:In biology, proteins are produced by
7:
339:The number of methods available for
190:The amino acids can be separated by
1202:Freeze-fracture electron microscopy
819:The Journal of Biological Chemistry
525:Identification by mass spectrometry
329:high-pressure liquid chromatography
178:evolved to determine the extent of
900:10.1111/j.1432-1033.1967.tb00047.x
552:The POI is isolated, typically by
453:onto a solid surface. One common
406:Separate and purify the fragments.
91:Determining amino acid composition
25:
700:Predicting from DNA/RNA sequences
315:) and dansyl derivatives such as
1182:Isothermal titration calorimetry
1162:Dual-polarization interferometry
887:European Journal of Biochemistry
653:Post-translational modifications
437:or by chemical reagents such as
420:Digestion into peptide fragments
55:post-translational modifications
449:The peptide to be sequenced is
881:Edman P, Begg G (March 1967).
594:collision-induced dissociation
335:C-terminal amino acid analysis
1:
1172:Chromatin immunoprecipitation
832:10.1016/S0021-9258(17)31291-7
241:-terminal amino acid analysis
1235:Protein structural alignment
1220:Protein structure prediction
979:10.1002/0471142727.mb1025s88
751:Applications to cryptography
1319:Super-resolution microscopy
1225:Protein function prediction
1153:Peptide mass fingerprinting
1148:Protein immunoprecipitation
579:Peptide mass fingerprinting
457:is glass fibre coated with
313:1-fluoro-2,4-dinitrobenzene
192:ion-exchange chromatography
186:Separation and quantitation
1374:
737:de novo peptide sequencing
535:de novo peptide sequencing
528:
484:acid. The derivative then
355:
1177:Surface plasmon resonance
1167:Microscale thermophoresis
1157:Protein mass spectrometry
1119:Green fluorescent protein
973:. Chapter 10: Unit10.25.
531:protein mass spectrometry
325:thin-layer chromatography
1197:Cryo-electron microscopy
674:Whole-mass determination
331:should be used instead.
216:ortho-phthaldehyde (OPA)
1230:Protein–protein docking
1143:Protein electrophoresis
1129:Protein immunostaining
936:10.1038/nprot.2006.468
883:"A protein sequenator"
500:
488:to give a substituted
375:in the protein with a
299:Hydrolyse the protein.
266:
100:
35:
1187:X-ray crystallography
498:
465:. The Edman reagent,
248:
98:
33:
1358:Proteomic sequencing
1114:Protein purification
728:Bioinformatics tools
467:phenylisothiocyanate
321:Phenylisothiocyanate
45:of all or part of a
1139:Gel electrophoresis
583:reversed phase HPLC
544:Proteolytic digests
490:phenylthiohydantoin
257:-terminal end with
196:reversed phase HPLC
110:amino acid analysis
43:amino acid sequence
1282:Display techniques
1134:Protein sequencing
634:post-translational
621:De novo sequencing
510:protein sequenator
501:
267:
253:derivatization of
207:isoelectric points
101:
78:protein sequenator
39:Protein sequencing
36:
1340:
1339:
1289:Bacterial display
867:978-1-61779-444-5
741:sequence analysis
694:mass spectrometry
598:post-source decay
504:Protein sequencer
381:2-mercaptoethanol
373:disulfide bridges
364:Edman degradation
358:Edman degradation
352:Edman degradation
345:carboxypeptidases
282:Edman degradation
230:and an optimised
214:reagents such as
133:hydrochloric acid
74:Edman degradation
70:mass spectrometry
18:Protein sequencer
16:(Redirected from
1365:
1304:Ribosome display
1240:Protein ontology
1086:
1079:
1072:
1063:
1057:
1011:
1010:
1000:
962:
956:
955:
924:Nature Protocols
919:
913:
912:
902:
878:
872:
871:
860:. Humana Press.
851:
845:
844:
834:
810:
722:molecular clones
640:N- and C-termini
463:cationic polymer
439:cyanogen bromide
385:protecting group
309:Sanger's reagent
259:Sanger's reagent
180:amide hydrolysis
21:
1373:
1372:
1368:
1367:
1366:
1364:
1363:
1362:
1343:
1342:
1341:
1336:
1313:
1277:
1273:Secretion assay
1249:
1206:
1100:
1090:
1060:
1038:10.1038/nrm1468
1023:
1019:
1017:Further reading
1014:
989:
964:
963:
959:
921:
920:
916:
880:
879:
875:
868:
853:
852:
848:
812:
811:
807:
803:
766:
753:
730:
702:
685:
676:
660:phosphorylation
655:
642:
623:
546:
537:
529:Main articles:
527:
506:
471:buffer solution
447:
422:
389:iodoacetic acid
360:
354:
337:
317:dansyl chloride
243:
188:
126:
93:
28:
23:
22:
15:
12:
11:
5:
1371:
1369:
1361:
1360:
1355:
1345:
1344:
1338:
1337:
1335:
1334:
1329:
1323:
1321:
1315:
1314:
1312:
1311:
1306:
1301:
1296:
1291:
1285:
1283:
1279:
1278:
1276:
1275:
1270:
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1259:
1257:
1251:
1250:
1248:
1247:
1242:
1237:
1232:
1227:
1222:
1216:
1214:
1212:Bioinformatics
1208:
1207:
1205:
1204:
1199:
1194:
1189:
1184:
1179:
1174:
1169:
1164:
1159:
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1145:
1136:
1131:
1126:
1121:
1116:
1110:
1108:
1102:
1101:
1091:
1089:
1088:
1081:
1074:
1066:
1059:
1058:
1032:(9): 699–711.
1020:
1018:
1015:
1013:
1012:
988:978-0471142720
987:
957:
930:(6): 2856–60.
914:
873:
866:
846:
804:
802:
799:
798:
797:
792:
787:
785:Donald F. Hunt
782:
777:
775:DNA sequencing
772:
765:
762:
752:
749:
733:Bioinformatics
729:
726:
701:
698:
684:
681:
675:
672:
654:
651:
641:
638:
622:
619:
618:
617:
613:
609:
601:
590:
571:
564:
561:
558:chromatography
545:
542:
526:
523:
505:
502:
475:trimethylamine
446:
443:
427:endopeptidases
421:
418:
417:
416:
413:
410:
407:
404:
401:
398:
395:
392:
377:reducing agent
356:Main article:
353:
350:
336:
333:
304:
303:
300:
297:
242:
236:
187:
184:
125:
122:
121:
120:
117:
92:
89:
26:
24:
14:
13:
10:
9:
6:
4:
3:
2:
1370:
1359:
1356:
1354:
1351:
1350:
1348:
1333:
1330:
1328:
1325:
1324:
1322:
1320:
1316:
1310:
1309:Yeast display
1307:
1305:
1302:
1300:
1299:Phage display
1297:
1295:
1292:
1290:
1287:
1286:
1284:
1280:
1274:
1271:
1269:
1268:Protein assay
1266:
1264:
1261:
1260:
1258:
1256:
1252:
1246:
1243:
1241:
1238:
1236:
1233:
1231:
1228:
1226:
1223:
1221:
1218:
1217:
1215:
1213:
1209:
1203:
1200:
1198:
1195:
1193:
1190:
1188:
1185:
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1178:
1175:
1173:
1170:
1168:
1165:
1163:
1160:
1158:
1154:
1151:
1149:
1146:
1144:
1140:
1137:
1135:
1132:
1130:
1127:
1125:
1122:
1120:
1117:
1115:
1112:
1111:
1109:
1107:
1103:
1098:
1094:
1087:
1082:
1080:
1075:
1073:
1068:
1067:
1064:
1055:
1051:
1047:
1043:
1039:
1035:
1031:
1027:
1022:
1021:
1016:
1008:
1004:
999:
994:
990:
984:
980:
976:
972:
968:
961:
958:
953:
949:
945:
941:
937:
933:
929:
925:
918:
915:
910:
906:
901:
896:
892:
888:
884:
877:
874:
869:
863:
859:
858:
850:
847:
842:
838:
833:
828:
824:
820:
816:
809:
806:
800:
796:
795:John R. Yates
793:
791:
790:Matthias Mann
788:
786:
783:
781:
780:Klaus Biemann
778:
776:
773:
771:
768:
767:
763:
761:
758:
750:
748:
746:
742:
738:
734:
727:
725:
723:
718:
716:
711:
710:transcription
707:
699:
697:
695:
689:
682:
680:
673:
671:
669:
665:
661:
652:
650:
648:
639:
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635:
631:
629:
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602:
599:
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584:
580:
576:
572:
569:
565:
562:
559:
555:
551:
550:
549:
543:
541:
536:
532:
524:
522:
520:
516:
511:
503:
497:
493:
491:
487:
483:
478:
476:
472:
468:
464:
460:
456:
452:
444:
442:
440:
436:
432:
428:
419:
414:
411:
408:
405:
402:
399:
396:
393:
390:
386:
382:
378:
374:
370:
369:
368:
365:
359:
351:
349:
346:
342:
334:
332:
330:
326:
322:
318:
314:
310:
301:
298:
295:
294:
293:
291:
287:
283:
279:
275:
273:
264:
260:
256:
252:
247:
240:
237:
235:
233:
229:
228:silica column
223:
221:
220:fluorescamine
217:
212:
208:
204:
199:
197:
193:
185:
183:
181:
177:
173:
169:
166:
162:
158:
154:
150:
146:
142:
138:
134:
130:
123:
118:
115:
114:
113:
111:
107:
97:
90:
88:
86:
84:
79:
75:
71:
66:
64:
60:
56:
52:
48:
44:
40:
32:
19:
1353:Cell biology
1294:mRNA display
1263:Enzyme assay
1133:
1124:Western blot
1106:Experimental
1029:
1025:
970:
960:
927:
923:
917:
893:(1): 80–91.
890:
886:
876:
856:
849:
825:(1): 531–9.
822:
818:
808:
754:
731:
719:
703:
690:
686:
677:
656:
646:
643:
627:
624:
547:
538:
514:
509:
507:
479:
448:
423:
361:
338:
308:
305:
289:
285:
271:
268:
262:
254:
250:
238:
224:
200:
189:
127:
109:
102:
82:
67:
38:
37:
1332:Vertico SMI
1192:Protein NMR
706:translation
683:Limitations
612:identified.
519:polypeptide
137:hydrophobic
59:translation
1347:Categories
801:References
770:Proteomics
630:sequencing
606:homologous
486:isomerises
371:Break any
341:C-terminal
153:tryptophan
129:Hydrolysis
124:Hydrolysis
596:(CID) or
575:MALDI-TOF
482:anhydrous
459:polybrene
455:substrate
274:-terminus
211:ninhydrin
157:glutamine
145:threonine
85:-terminus
1099:of study
1093:Proteins
1046:15340378
1007:19816929
944:17406544
764:See also
757:proposed
554:SDS-PAGE
451:adsorbed
445:Reaction
429:such as
387:such as
261:(DNFB),
168:reagents
161:cysteine
149:tyrosine
106:protease
76:using a
1097:methods
1054:5176895
998:2905857
952:8248224
909:6059350
841:2642478
715:UniProt
647:de novo
628:de novo
568:Trypsin
473:of 12%
431:trypsin
278:peptide
232:elution
176:ammonia
51:peptide
47:protein
1095:: key
1052:
1044:
1005:
995:
985:
950:
942:
907:
864:
839:
435:pepsin
172:phenol
159:, and
141:serine
1255:Assay
1050:S2CID
948:S2CID
745:BLAST
379:like
276:of a
165:thiol
63:genes
1042:PMID
1003:PMID
983:ISBN
940:PMID
905:PMID
862:ISBN
837:PMID
533:and
461:, a
383:. A
362:The
72:and
1034:doi
993:PMC
975:doi
932:doi
895:doi
827:doi
823:264
747:).
713:as
668:ECD
666:or
664:ETD
587:ESI
556:or
433:or
327:or
218:or
170:or
61:of
49:or
1349::
1048:.
1040:.
1028:.
1001:.
991:.
981:.
969:.
946:.
938:.
926:.
903:.
889:.
885:.
835:.
821:.
817:.
696:.
508:A
319:.
222:.
203:pH
198:.
182:.
155:,
151:,
147:,
143:,
87:.
65:.
1155:/
1141:/
1085:e
1078:t
1071:v
1056:.
1036::
1030:5
1009:.
977::
954:.
934::
928:1
911:.
897::
891:1
870:.
843:.
829::
560:.
515:N
311:(
290:N
286:N
272:N
263:B
255:N
251:A
239:N
83:N
20:)
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