Characterization and localization of protein modifications
A method for characterizing proteins, including steps of (a) detecting a plurality of proteins, wherein individual proteins of the plurality are associated with unique identifiers, wherein the detecting distinguishes the identities of the individual proteins and the unique identifiers associated with the individual proteins; (b) digesting the proteins to form peptides, wherein the peptides from each protein are associated with the unique identifiers for the respective individual protein; (c) detecting the peptides and associated unique identifiers, wherein the detecting distinguishes characteristics of individual peptides, and wherein the detecting distinguishes unique identifiers associated with the individual peptides; and (d) correlating characteristics detected in step (c) with individual proteins detected in step (a) based on the unique identifiers associated with the individual proteins and the peptides.
1 . A composition, comprising:
a) a nucleic acid particle; and
b) a polypeptide attached to the nucleic acid particle through two or more linking moieties,
wherein first ends of the two or more linking moieties are attached to the nucleic acid particle and second ends of the two or more linking moieties are attached to the polypeptide.
2 . The composition of claim 1 , wherein the polypeptide comprises a full-length protein.
3 . The composition of claim 1 , wherein the composition comprises a plurality of polypeptides, and each polypeptide in the plurality of polypeptides is attached to two or more linking moieties.
4 . The composition of claim 1 , wherein each of the two or more linking moieties comprises a same identifier label.
5 . The composition of claim 1 , wherein the polypeptide is attached to a linking moiety of the two or more linking moieties by a bond between the polypeptide and an identifier label attached to the linking moiety.
6 . The composition of claim 1 , wherein the polypeptide comprises at least one cleavage site.
7 . The composition of claim 6 , wherein the cleavage site is located on the polypeptide between an attachment site of a first linking moiety of the two or more linking moieties and a second attachment site of a second linking moiety of the two or more linking moieties.
8 . The composition of claim 1 , wherein the polypeptide comprises a post-translational modification.
9 . The composition of claim 8 , wherein the composition comprises a plurality of polypeptides and a second polypeptide in the composition does not comprise the post-translational modification.
10 . The composition of claim 1 , further comprising a solid support.
11 . The composition of claim 10 , wherein the nucleic acid particle is attached to the solid support.
12 . The composition of claim 11 , wherein the nucleic acid particle is contained within a well of the solid support.
13 . The composition of claim 12 , wherein the nucleic acid particle is attached to a bead, and wherein the bead is contained in the well of the solid support.
14 . The composition of claim 1 , further comprising an affinity reagent bound to the polypeptide.
15 . The composition of claim 14 , wherein the affinity reagent is bound to a post-translational modification of the polypeptide.
16 . The composition of claim 14 , wherein the affinity reagent is bound to a trimer, tetramer, pentamer, or hexamer epitope of the polypeptide.
17 . The composition of claim 1 , wherein the polypeptide further comprises a molecular barcode.
18 . An array comprising a first address for binding a first polypeptide and a second address for binding a second polypeptide, wherein the first address and the second address are optically resolvable, wherein the first address is attached to a first nucleic acid particle, wherein the second address is attached to a second nucleic acid particle, and wherein:
a) the first nucleic acid particle comprises two or more linking moieties, wherein the two or more linking moieties of the first nucleic acid particle are attached to the first polypeptide; and
b) the second nucleic acid particle comprises two or more linking moieties, wherein the two or more linking moieties of the second nucleic acid particle are attached to the second polypeptide;
wherein the first polypeptide differs from the second polypeptide.
19 . A system comprising:
a) a solid support, wherein an address of the solid support is attached to a nucleic acid particle, wherein the particle comprises two or more linking moieties, wherein the two or more linking moieties of the nucleic acid particle are attached to a polypeptide;
b) a detectable affinity reagent, wherein the detectable affinity reagent is configured to provide a detectable signal, and wherein the detectable affinity reagent is configured to be delivered to the solid support;
c) an optical detector; and
d) one or more processors programmed to read data from the optical detector to detect a presence or absence of the detectable signal at the address of the solid support.