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Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture Agricultural Research Service Western Regional Research Center Albany, California Gluten Proteins in Relation to Celiac Disease

Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

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Gluten Proteins in Relation to Celiac Disease. Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture Agricultural Research Service Western Regional Research Center Albany, California. Diagram of a Wheat Kernel. starch granule. endosperm protein. - PowerPoint PPT Presentation

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Page 1: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

Donald D. Kasarda, Ph.D.

Research Chemist (Retired)/Collaborator

U.S. Department of Agriculture

Agricultural Research Service

Western Regional Research Center

Albany, California

Gluten Proteins in Relation to

Celiac Disease

Page 2: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 3: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

Diagram of aWheat Kernel

Page 4: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

FLOUR PARTICLES

starch granule

endosperm protein

Page 5: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 6: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 7: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

Glutamine/Proline Composition of Wheat Prolamins (Mole Percent)

gliadins

gliadins

LMW-glutenin subunits

HMW-glutenin subunits

Gln Pro Gln + Pro

43-54 20-29 63-83

36-42 15-19 51-61+

30-38 12-27 42-55

Page 8: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 9: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

2-Dimensional electrophoretic pattern of gluten proteins

Page 10: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 11: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 12: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

“Consensus” Repeat Types for Gluten Proteins

-P-Q-Q-P-(Y/F)-P-gliadins

-gliadins -Q-Q-P-Q-Q-P-F-P-

D-type -gliadins

B-type -gliadins -Q-Q-Q-F-P-

-Q-P-Q-Q-P-F-P-

LMW-glutenin subunits

HMW-glutenin subnits

-Q-Q-Q-Q-P-P-F-S-

-Q-Q-G-Y-Y-P-T-S-P-

Page 13: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 14: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 15: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

Leu

GlyGln Gln Gln

Gln GlnPro Pro Pro

ProTyr

Phe

Molecular model of a 13-residue gliadin peptide (a 13-mer)

L-G-Q-Q-Q-P-FF-P-P-Q-Q-P-Y

Page 16: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 17: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 18: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 19: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture
Page 20: Donald D. Kasarda, Ph.D. Research Chemist (Retired)/Collaborator U.S. Department of Agriculture

Currently Favored Analytical Method for Gluten in Food

R-5 monoclonal antibody ELISA test is available commercially.

Detection down to about 1.5 parts per million; quantification above 2.5 parts per million has been reported.

Antibody reacts with monomeric wheat, rye, and barley prolamins, butnot oat avenins; reacts weakly or not at all with glutelins (glutenin).

The Codex Alimentarius Committee on Methods agreed in 2004 to endorse temporarily the R5 ELISA for the determination of gluten.

Possible problems: failure to detect glutelin (glutenin) proteins andsmall peptides from hydrolysates.

Recognizes QQPFP and LQPFP motifs most strongly.