title: Development of soluble ester-linked aldehyde polymers for proteomics.
abstract: High molecular weight hyperbranched polyglycerol (HPG) was selected for development as a soluble polymer support for the targeted selection and release of primary-amine containing peptides from a complex mixture. HPG has been functionalized with ester-linked aldehyde groups that can bind primary-amine containing peptides via a reductive alkylation reaction. Once bound, the high molecular weight of the polymer facilitates separation from a complex peptide mixture by employing either a 30 kDa molecular weight cutoff membrane or precipitation in acetonitrile. Following the removal of unbound peptides and reagents, subsequent hydrolysis of the ester linker releases the bound peptide into solution for analysis by mass spectrometry. Released peptides retain the linker moiety and are therefore characteristically mass-shifted. Four water-soluble cleavable aldehyde polymers (CAP1, CAP2, CAP3, and CAP4) ranging in types of linker groups, length of the linker groups, have been prepared and characterized, each demonstrating the ability to selectively enrich and sequence primary-amine peptides from a complex human proteome containing blocked (dimethylated amine) and unblocked (primary amine) peptides. The polymers have very low nonspecific peptide-binding properties while possessing significantly more reactive groups per milligram of the support than commercially available resins. The polymers exhibit a range of reactivities and binding capacities that depend on the type of linker group between the aldehyde group and the polymer. Using various linker structures, we also probed the mechanism of the observed dehydration of hydrolyzed peptides during matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis.
authors.affiliation: Centre for Blood Research, The University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors.forename: Patrick
authors.initials: P
authors.lastname: Beaudette
authors.forename: Nicholas A A
authors.initials: NA
authors.lastname: Rossi
authors.forename: Pitter F
authors.initials: PF
authors.lastname: Huesgen
authors.forename: Xifei
authors.initials: X
authors.lastname: Yu
authors.forename: Rajesh A
authors.initials: RA
authors.lastname: Shenoi
authors.forename: Alain
authors.initials: A
authors.lastname: Doucet
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Overall
authors.forename: Jayachandran N
authors.initials: JN
authors.lastname: Kizhakkedathu
mesh_terms: D000447:Aldehydes / Q000737:chemistry*; D004952:Esters; D005990:Glycerol / Q000138:chemical synthesis / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000447:Aldehydes; D004952:Esters; D010455:Peptides; D011108:Polymers; C043941:polyglycerol; D005990:Glycerol