title: Development of Diagnostic Fragment Ion Library for Glycated Peptides of Human Serum Albumin: Targeted Quantification in Prediabetic, Diabetic, and Microalbuminuria Plasma by Parallel Reaction Monitoring, SWATH, and MSE.
abstract: Human serum albumin is one of the most abundant plasma proteins that readily undergoes glycation, thus glycated albumin has been suggested as an additional marker for monitoring glycemic status. Hitherto, only Amadori-modified peptides of albumin were quantified. In this study, we report the construction of fragment ion library for Amadori-modified lysine (AML), N(ε)-(carboxymethyl)lysine (CML)-, and N(ε)-(carboxyethyl)lysine (CEL)-modified peptides of the corresponding synthetically modified albumin using high resolution accurate mass spectrometry (HR/AM). The glycated peptides were manually inspected and validated for their modification. Further, the fragment ion library was used for quantification of glycated peptides of albumin in the context of diabetes. Targeted Sequential Window Acquisition of all THeoretical Mass Spectra (SWATH) analysis in pooled plasma samples of control, prediabetes, diabetes, and microalbuminuria, has led to identification and quantification of 13 glycated peptides comprised of four AML, seven CML, and two CEL modifications, representing nine lysine sites of albumin. Five lysine sites namely K549, K438, K490, K88, and K375, were observed to be highly sensitive for glycation modification as their respective m/z showed maximum fold change and had both AML and CML modifications. Thus, peptides involving these lysine sites could be potential novel markers to assess the degree of glycation in diabetes.
authors.affiliation: From the §Mass-Spectrometry and Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune-411008, India.;
authors.forename: Arvind M
authors.initials: AM
authors.lastname: Korwar
authors.affiliation: From the §Mass-Spectrometry and Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune-411008, India.;
authors.forename: Garikapati
authors.initials: G
authors.lastname: Vannuruswamy
authors.affiliation: From the §Mass-Spectrometry and Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune-411008, India.;
authors.forename: Mashanipalya G
authors.initials: MG
authors.lastname: Jagadeeshaprasad
authors.affiliation: From the §Mass-Spectrometry and Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune-411008, India.;
authors.forename: Ramesha H
authors.initials: RH
authors.lastname: Jayaramaiah
authors.affiliation: From the §Mass-Spectrometry and Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune-411008, India.;
authors.forename: Shweta
authors.initials: S
authors.lastname: Bhat
authors.affiliation: ‡Madras Diabetes Research Foundation, Chennai- 600 086, India.
authors.forename: Bhaskaran S
authors.initials: BS
authors.lastname: Regin
authors.affiliation: From the §Mass-Spectrometry and Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune-411008, India.;
authors.forename: Sureshkumar
authors.initials: S
authors.lastname: Ramaswamy
authors.affiliation: From the §Mass-Spectrometry and Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune-411008, India.;
authors.forename: Ashok P
authors.initials: AP
authors.lastname: Giri
authors.affiliation: ‡Madras Diabetes Research Foundation, Chennai- 600 086, India.
authors.forename: Viswanathan
authors.initials: V
authors.lastname: Mohan
authors.affiliation: ‡Madras Diabetes Research Foundation, Chennai- 600 086, India.
authors.forename: Muthuswamy
authors.initials: M
authors.lastname: Balasubramanyam
authors.affiliation: From the §Mass-Spectrometry and Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune-411008, India.; mj.kulkarni@ncl.res.in.
authors.forename: Mahesh J
authors.initials: MJ
authors.lastname: Kulkarni
mesh_terms: D000419:Albuminuria / Q000097:blood / Q000378:metabolism*; D000595:Amino Acid Sequence; D000704:Analysis of Variance; D003920:Diabetes Mellitus / Q000097:blood / Q000378:metabolism*; D017127:Glycation End Products, Advanced; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives / Q000378:metabolism; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011236:Prediabetic State / Q000378:metabolism*; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000093423:Glycated Serum Albumin
chemical_list: D017127:Glycation End Products, Advanced; D019151:Peptide Library; D010455:Peptides; D012709:Serum Albumin; C048496:N(6)-carboxymethyllysine; D008239:Lysine; D000093423:Glycated Serum Albumin