Summary.
We report an investigation of the site specificity, extent and nature of modification of bovine serum albumin (BSA) incubated with fructose or glucose at physiological temperature and pH. Sites of early glycation (Heyns rearrangement products (HRP) from fructose; fructoselysine (FL) from glucose) as well as advanced glycation (N ε-(carboxymethyl)lysine; CML) were analyzed by liquid chromatography-mass spectrometry. The major site of modification by fructose, like glucose, is Lysine-524 and this results in, respectively, 31 and 76% loss of the corresponding unmodified tryptic peptide, Gln525-Lys533. In addition, total lysine, HRP, FL, CML and N ε-(carboxyethyl)lysine in the incubations, was quantified. Almost all of the loss of lysine in the fructose-modified BSA was attributed to the formation of CML, with the yield of CML being up to 17-fold higher than glucose-modified BSA. A mechanism for the formation of CML from the HRP is proposed.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
MU Ahmed SR Thorpe JW Baynes (1986) ArticleTitleIdentification of N-epsilon-(carboxymethyl)lysine as a degradation product of fructoselysine in glycated protein J Biol Chem 261 4889–4894 Occurrence Handle3082871 Occurrence Handle1:CAS:528:DyaL28Xkt1anu74%3D
MU Ahmed E Brinkmann-Frye TP Degenhardt SR Thorpe JW Baynes (1997) ArticleTitleNε-(Carboxyethyl)lysine, a product of the chemical modification of proteins by methylglyoxal, increases with age in humen lens proteins Biochem J 324 565–570 Occurrence Handle9182719 Occurrence Handle1:CAS:528:DyaK2sXjvFGmtLc%3D
MAM van Boekel HJ Hoenders (1992) ArticleTitleGlycation of crystallins in lenes from aging and diabetic individuals FEBS Lett 314 1–4 Occurrence Handle1451795 Occurrence Handle1:CAS:528:DyaK3sXjslSgsg%3D%3D Occurrence Handle10.1016/0014-5793(92)81446-S
JWC Brock DJS Hinton WE Cotham TO Metz SR Thorpe JW Baynes JM Ames (2003) ArticleTitleProteomic analysis of the site specificity of glycation and carboxymethylation of ribonuclease J Proteome Res 2 506–513 Occurrence Handle14582647 Occurrence Handle1:CAS:528:DC%2BD3sXkvFyjs7k%3D Occurrence Handle10.1021/pr0340173
EB Casey HR Zhao EC Abraham (1995) ArticleTitleRole of glycine 1 and Lysine 2 in the glycation of bovine γB-crystallin J Biol Chem 270 20781–20786 Occurrence Handle7657661 Occurrence Handle1:CAS:528:DyaK2MXnvVOqu78%3D Occurrence Handle10.1074/jbc.270.39.23119
MP Cohen (2003) ArticleTitleIntervention strategies to prevent pathogenic effects of glycated albumin Arch Biochem Biophys 419 25–30 Occurrence Handle14568005 Occurrence Handle1:CAS:528:DC%2BD3sXotVCjur8%3D Occurrence Handle10.1016/j.abb.2003.08.012
WE Cotham TO Metz PL Ferguson JWC Brock DJS Hinton SR Thorpe JW Baynes JM Ames (2004) ArticleTitleProteomic analysis of arginine adducts on glyoxal-modified ribonuclease Mol Cell Proteomics 3 1145–1153 Occurrence Handle15377717 Occurrence Handle1:CAS:528:DC%2BD2MXhvFarsQ%3D%3D Occurrence Handle10.1074/mcp.M400002-MCP200
JA Dunn JS Patrick SR Thorpe JW Baynes (1989) ArticleTitleOxidation of glycated proteins: age-dependent accumulation of Nε-(carboxymethyl)lysine in lens proteins Biochemistry 28 9464–9468 Occurrence Handle2514802 Occurrence Handle1:CAS:528:DyaL1MXmt12ls7k%3D Occurrence Handle10.1021/bi00450a033
JA Dunn DR McCance SR Thorpe TJ Lyons JW Baynes (1991) ArticleTitleAge-dependent accumulation of Nε-(carboxymethyl)lysine and Nε-(carboxymethyl)hydroxylysine in human skin collagen Biochemistry 30 1205–1210 Occurrence Handle1899338 Occurrence Handle1:CAS:528:DyaK3MXlvFCmug%3D%3D Occurrence Handle10.1021/bi00219a007
KH Gabbay JH Kinoshita (1972) ArticleTitleMechanism of development and possible prevention of sugar cataracts Isr J Med Sci 8 1557–1561 Occurrence Handle4647820 Occurrence Handle1:CAS:528:DyaE3sXisFem
RL Garlick JS Mazer (1983) ArticleTitleThe principal site of nonenzymatic glycosylation of human serum albumin in vivo J Biol Chem 258 6142–6146 Occurrence Handle6853480 Occurrence Handle1:CAS:528:DyaL3sXktlCksrY%3D
MA Glomb VM Monnier (1995) ArticleTitleMechanism of protein modification by glyoxal and glycoaldehyde, reactive intermediates of the Maillard reaction J Biol Chem 270 10017–10026 Occurrence Handle7730303 Occurrence Handle1:CAS:528:DyaK2MXltlWnsrY%3D Occurrence Handle10.1074/jbc.270.17.10017
DJ Heaf DJ Galton (1975) ArticleTitleSorbitol and other polyols in lens, adipose tissue and urine in diabetes mellitus Clin Chim Acta 63 41–47 Occurrence Handle1181073 Occurrence Handle1:STN:280:DyaE28%2Fkt1ehsA%3D%3D Occurrence Handle10.1016/0009-8981(75)90376-9
HG Hers (1956) ArticleTitleLe mécanisme de la transformation de glucose en fructose par les vésicules séminales Biochim Biophys Acta 22 202–203 Occurrence Handle13373872 Occurrence Handle1:CAS:528:DyaG2sXhsVWrug%3D%3D Occurrence Handle10.1016/0006-3002(56)90247-5
K Heyns G Müller H Paulsen (1967) ArticleTitleQuantitative investigation of the reaction of hexoses with amino acids Justus Liebigs Ann Chem 703 202–214 Occurrence Handle5593875 Occurrence Handle1:CAS:528:DyaF2sXkvFWisr4%3D
N Iberg R Flückinger (1986) ArticleTitleNonenzymic glycosylation of albumin in vivo: Identification of multiple glycosylation sites J Biol Chem 261 13542–13545 Occurrence Handle3759977 Occurrence Handle1:CAS:528:DyaL28XlvVKhsL0%3D
JA Jedziniak LT Chylack SuffixJr HM Cheng MK Gillis AA Kalustian WH Tung (1981) ArticleTitleThe sorbitol pathway in the human lens: aldose reductase and polyol dehydrogenase Invest Ophthalmol Vis Sci 20 314–326 Occurrence Handle6782033 Occurrence Handle1:CAS:528:DyaL3MXkslOlt7Y%3D
A Lapolla D Fedele R Reitano NC Aricò R Seraglia P Traldi E Marotta R Tonani (2004) ArticleTitleEnzymatic digestion and mass spectrometry in the study of advanced glycation end products/peptides Am J Mass Spectrom 15 496–509 Occurrence Handle1:CAS:528:DC%2BD2cXisVGhurs%3D Occurrence Handle10.1016/j.jasms.2003.11.014
J Leonil D Molle J Fauquant JL Maubois RJ Pearce S Bouhallab (1997) ArticleTitleCharacterization by ionisation mass spectrometry of lactosyl β-lactoglobulin conjugates formed during heat treatment of milk and whey and identification of one lactose-binding site J Dairy Sci 80 2270–2281 Occurrence Handle9361199 Occurrence Handle1:CAS:528:DyaK2sXntFems74%3D Occurrence Handle10.3168/jds.S0022-0302(97)76176-9
I Macdonald A Keyser D Pacy (1978) ArticleTitleSome effects in man, of varying the load of glucose, sucrose, fructose or sorbitol on metabolites in blood Am J Clin Nutr 31 1305–1311 Occurrence Handle677070 Occurrence Handle1:CAS:528:DyaE1cXlsVGmsb0%3D
H Makino K Shikata M Kushiro K Hironaka Y Yamasaki H Sugimoto Z Ota N Araki S Horiuchi (1996) ArticleTitleRoles of advanced glycation end-products in the progression of diabetic nephropathy Nephrol Dial Transpl 11 76–80 Occurrence Handle1:CAS:528:DyaK2sXhsFCitbc%3D
E Marotta A Lapolla D Fedele A Senesi R Reitano M Witt R Seraglia P Traldi (2003) ArticleTitleAccurate mass measurements by Fourier transform mass spectrometry in the study of advanced glycation end products/peptides J Mass Spectrom 38 196–205 Occurrence Handle12577286 Occurrence Handle1:CAS:528:DC%2BD3sXhsFGmurk%3D Occurrence Handle10.1002/jms.431
JD McPherson BH Shilton DJ Walton (1988) ArticleTitleRole of fructose in glycation and cross-linking of proteins Biochemistry 27 1901–1907 Occurrence Handle3132203 Occurrence Handle1:CAS:528:DyaL1cXht1Kqurs%3D Occurrence Handle10.1021/bi00406a016
Y Nakamura Y Horii T Nishino H Shiiki Y Sakaguchi T Kagoshima K Dohi Z Makita H Vlassara R Bucula (1993) ArticleTitleImmunohistochemical localization of advanced glycosylation endproducts in coronary atheroma and cardiac tissue in diabetes-mellitus Am J Pathol 143 1649–1656 Occurrence Handle8256853 Occurrence Handle1:STN:280:DyaK2c%2FnvFSnsA%3D%3D
BJ Ortwerth SH Slight M Prabhakaram Y Sun JB Smith (1992) ArticleTitleSite-specific glycation of lens crystallins by ascorbic acid Biochim Biophys Acta 1117 207–215 Occurrence Handle1525182 Occurrence Handle1:CAS:528:DyaK38XmtlWmsL4%3D
P Pam SA McLuckey (2003) ArticleTitleElectrospray ionization of protein mixtures at low pH Anal Chem 75 1491–1499 Occurrence Handle10.1021/ac020637w Occurrence Handle1:CAS:528:DC%2BD3sXht1SgsLo%3D
J Pennington JJ Harding (1994) ArticleTitleIdentification of the site of glycation of γ-II-crystallin by (14C)-fructose Biochim Biophys Acta 1226 163–167 Occurrence Handle8204663 Occurrence Handle1:CAS:528:DyaK2cXktlagur0%3D
S Reddy J Bichler KJ Wells-Knecht SR Thorpe J Baynes (1995) ArticleTitleN-Epsilon-(carboxymethyl)lysine is a dominant advanced glycation endproduct (AGE) antigen in tissue proteins Biochemistry 34 10872–10878 Occurrence Handle7662668 Occurrence Handle1:CAS:528:DyaK2MXnsVKrtbY%3D Occurrence Handle10.1021/bi00034a021
A Ruttkat HF Ebersdobler (1995) ArticleTitleNε-Carboxymethyllysine is formed during heating of lysine with ketoses J Sci Food Agric 68 261–263 Occurrence Handle1:CAS:528:DyaK2MXmt12msLc%3D
M Sakai M Oimomi M Kasuga (2002) ArticleTitleExperimental studies on the role of fructose in the development of diabetic complications Kobe J Med Sci 48 125–136 Occurrence Handle12594356 Occurrence Handle1:CAS:528:DC%2BD3sXmtVClu7o%3D
P Saxena AK Saxena XL Cui M Obenovich K Gudipaty VM Monnier (2000) ArticleTitleTransition metal-catalyzed oxidation of ascorbate in human cataract extracts: possible role of advanced glycation end products Invest Opthalmol Vis Sci 41 1473–1481 Occurrence Handle1:STN:280:DC%2BD3c3ls1GhsQ%3D%3D
DB Shin F Hayase H Kato (1988) ArticleTitlePolymerization of proteins caused by reaction with sugars and the formation of 3-deoxyglucosone under physiological conditions Agric Biol Chem 52 1451–1458 Occurrence Handle1:CAS:528:DyaL1cXltVKlsLg%3D
MA Smith S Taneda PL Richey S Miyata S-D Yan D Stern LM Sayre VM Monnier G Perry (1994) ArticleTitleAdvanced Maillard reaction end products are associated with Alzheimer’s disease pathology Proc Natl Acad Sci USA 91 5710–5714 Occurrence Handle8202552 Occurrence Handle1:CAS:528:DyaK2cXksFGisb4%3D Occurrence Handle10.1073/pnas.91.12.5710
G Suarez R Rajaram AL Oronsky MA Gawinowicz (1989) ArticleTitleNonenzymic glycation of bovine serum albumin by fructose (fructation). Comparison with the Maillard reaction initiated by glucose J Biol Chem 264 3674–3679 Occurrence Handle2537288 Occurrence Handle1:CAS:528:DyaL1MXhsFSisrY%3D
PJ Thornalley A Langborg HS Minhas (1999) ArticleTitleFormation of glyoxal, methylglyoxal and 3-deoxyglucosone in the glycation of proteins by glucose Biochem J 344 109–116 Occurrence Handle10548540 Occurrence Handle1:CAS:528:DyaK1MXns1yjtrY%3D Occurrence Handle10.1042/0264-6021:3440109
JV Valencia SC Weldon D Quinn GH Kiers J DeGroot JM TeKoppele TE Hughes (2004) ArticleTitleAdvanced glycation end product ligands for the receptor for advanced glycation end products: biochemical characterization and formation kinetics Anal Biochem 324 68–78 Occurrence Handle14654047 Occurrence Handle1:CAS:528:DC%2BD3sXptl2qt74%3D Occurrence Handle10.1016/j.ab.2003.09.013
MC Wells-Knecht SR Thorpe JW Baynes (1995) ArticleTitlePathways of formation of glycoxidation products during glycation of collagen Biochemistry 34 15134–15141 Occurrence Handle7578127 Occurrence Handle1:CAS:528:DyaK2MXovFOju7o%3D Occurrence Handle10.1021/bi00046a020
FK Yeboah I Alli VA Yaylayan (1999) ArticleTitleReactivities of D-glucose and D-fructose during glycation of bovine serum albumin J Agric Food Chem 47 3164–3172 Occurrence Handle10552625 Occurrence Handle1:CAS:528:DyaK1MXksFGhsrw%3D Occurrence Handle10.1021/jf981289v
HR Zhao JB Smith XY Jiang EC Abraham (1996) ArticleTitleSites of glycation of beta B-2-crystallin by glucose and fructose Biochem Biophys Res Commun 229 128–133 Occurrence Handle8954094 Occurrence Handle1:CAS:528:DyaK28Xnt12rsb8%3D Occurrence Handle10.1006/bbrc.1996.1768
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hinton, D., Ames, J. Site specificity of glycation and carboxymethylation of bovine serum albumin by fructose. Amino Acids 30, 425–434 (2006). https://doi.org/10.1007/s00726-006-0269-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-006-0269-2