Table 1. Mass-barcoded peptide substrate sequences.
Table describing the sequences of the mass-barcoded peptide substrates along with their chemical modifications. ANP was used as a photocleavable linker to enable rapid detachment from the microparticles. 5-FAM was used for rapid quantification via fluorescence. Isotope enrichment modifications were used to distinguish mass barcodes for quantification with mass spectrometry.
| Substrate Name | Peptide sequence (N terminus on left)* | Modifications** |
|---|---|---|
| CC01 | e(*aa)(*aa)ndneeGFFsAr(ANP)K(5-FAM)GGLQRIYKC | 1st *aa = Gly(13C2); 2nd *aa = Val(U13C5,15N) |
| CC02 | eG(*aa)ndneeGF(*aa)s(*aa)r(ANP)K(5-FAM)GGKSVARTLLVKC | 1st *aa = Val(U13C5,15N); 2nd *aa = Phe(15N); 3rd *aa = Ala(15N) |
| CC03 | e(*aa)(*aa)ndneeGFFs(*aa)r(ANP)K(5-FAM)GGQRQRIIGGC | 1st *aa = Gly(U13C2,15N); 2nd *aa = Val(15N); 3rd *aa = Ala (U13C3,15N) |
| CC04 | e(*aa)Vndnee(*aa)FFs(*aa)r(ANP)K(5-FAM)GGKYLGRSYKVC | 1st *aa = Gly(13C2); 2nd *aa = Gly(13C2); 3rd *aa = Ala(U13C3,15N) |
| CC05 | eGVndnee(*aa)(*aa)Fs(*aa)r(ANP)K(5-FAM)GGGLQRALEIC | 1st *aa = Gly(U13C2,15N); 2nd *aa = Phe(15N); 3rd *aa = Ala(U13C3,15N) |
| CC06 | e(*aa)(*aa)ndnee(*aa)(*aa)(*aa)s(*aa)r(ANP)K(5-FAM)GGKTTGGRIYGGC | 1st *aa = Gly(13C2); 2nd *aa = Val(U13C5,15N); 3rd *aa = Gly(U13C2,15N); 4th *aa = Phe(15N); 5th *aa = Phe(15N); 6th *aa = Ala(15N); still include ANP and K5-FAM |
| CC07 | eG(*aa)ndnee(*aa)(*aa)Fs(*aa)r(ANP)K(5-FAM)GGQARGGSC | 1st *aa = Val(U13C5,15N); 2nd *aa = Gly(U13C2,15N); 3rd *aa = Phe(15N); 4th *aa = Ala(U13C3,15N) |
*ANP = Photocleavable linker 3-Amino-3-(2-nitrophenyl)propionic acid
*5-FAM = 5—Carboxyfluorescein
**Modifications represent heavy amino acids (i.e., isotope enrichment)