Streamlining Peptide Mapping LC-MS Approach for Studying Fusion Peptide-Conjugated Vaccine Immunogens

2021 ◽  
Author(s):  
Vera B. Ivleva ◽  
Daniel B. Gowetski ◽  
Q. Paula Lei
Keyword(s):  
Peptide Mapping ◽  
Fusion Peptide

Destabilization of a model membrane by a predicted fusion peptide of fertilin α

1998 ◽  
Vol 95 (2) ◽  
pp. 467-473 ◽  
Author(s):  
A. Schanck ◽  
R. Brasseur ◽  
J. Peuvot
Keyword(s):  
Fusion Peptide ◽  
Model Membrane

scholarly journals Peptide mapping of the human transferrin receptor in normal and transformed cells.

1983 ◽  
Vol 258 (4) ◽  
pp. 2668-2673
Author(s):  
B S Stein ◽  
H H Sussman
Keyword(s):  
Transferrin Receptor ◽  
Peptide Mapping ◽  
Transformed Cells ◽  
Human Transferrin ◽  
Human Transferrin Receptor

scholarly journals Deamidation and glycation of a Bacillus licheniformis α-amylase during industrial fermentation can improve detergent wash performance

Amylase ◽  
2021 ◽  
Vol 5 (1) ◽  
pp. 38-49
Author(s):  
Connie Pontoppidan ◽  
Svend G. Kaasgaard ◽  
Carsten P. Sønksen ◽  
Carsten Andersen ◽  
Birte Svensson
Keyword(s):  
Bacillus Licheniformis ◽  
Mutational Analysis ◽  
Peptide Mapping ◽  
Substrate Binding ◽  
Carbohydrate Binding ◽  
Surface Binding ◽  
Lysine Residues ◽  
Tandem Mass Spectrometric ◽  
Binding Cleft ◽  
Household Detergents

Abstract The industrial thermostable Bacillus licheniformis α-amylase (BLA) has wide applications, including in household detergents, and efforts to improve its performance are continuously ongoing. BLA during the industrial production is deamidated and glycated resulting in multiple forms with different isoelectric points. Forty modified positions were identified by tandem mass spectrometric peptide mapping of BLA forms separated by isoelectric focusing. These modified 12 asparagine, 9 glutamine, 8 arginine and 11 lysine residues are mostly situated on the enzyme surface and several belong to regions involved in stability, activity and carbohydrate binding. Eight residues presumed to interact with starch at the active site and surface binding sites (SBSs) were subjected to mutational analysis. Five mutants mimicking deamidation (N→D, Q→E) at the substrate binding cleft showed moderate to no effect on thermostability and k cat and K M for maltoheptaose and amylose. Notably, the mutations improved laundry wash efficiency in detergents at pH 8.5 and 10.0. Replacing three reducing sugar reactive side chains (K→M, R→L) at a distant substrate binding region and two SBSs enhanced wash performance especially in liquid detergent at pH 8.5, slightly improved enzymatic activity and maintained thermostability. Wash performance was most improved (5-fold) for the N265D mutant near substrate binding subsite +3.

scholarly journals The Development of Cleveland Peptide Mapping by Don W. Cleveland

2006 ◽  
Vol 281 (33) ◽  
pp. e27-e28
Author(s):  
Nicole Kresge ◽  
Robert D. Simoni ◽  
Robert L. Hill
Keyword(s):  
Peptide Mapping

scholarly journals Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis.

1977 ◽  
Vol 252 (3) ◽  
pp. 1102-1106 ◽  
Author(s):  
D W Cleveland ◽  
S G Fischer ◽  
M W Kirschner ◽  
U K Laemmli
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Peptide Mapping ◽  
Sodium Dodecyl ◽  
Limited Proteolysis ◽  
Dodecyl Sulfate

scholarly journals Purification and peptide mapping of rat brain choline acetyltransferase.

1980 ◽  
Vol 255 (22) ◽  
pp. 10612-10617
Author(s):  
G.W. Dietz ◽  
P.M. Salvaterra
Keyword(s):  
Rat Brain ◽  
Choline Acetyltransferase ◽  
Peptide Mapping

scholarly journals Characterization of a novel + 70 Da modification in rhGM-CSF expressed in E. coli using chemical assays in combination with mass spectrometry

Amino Acids ◽  
2021 ◽  
Author(s):  
Magdalena Widgren Sandberg ◽  
Jakob Bunkenborg ◽  
Stine Thyssen ◽  
Martin Villadsen ◽  
Thomas Kofoed
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Peptide Mapping ◽  
Covalent Modification ◽  
Peptide Fragmentation ◽  
Tandem Mass ◽  
E Coli ◽  
Gm Csf ◽  
Fragmentation Behavior ◽  
Macrophage Colony

AbstractGranulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine and a white blood cell growth factor that has found usage as a therapeutic protein. During analysis of different fermentation batches of GM-CSF recombinantly expressed in E. coli, a covalent modification was identified on the protein by intact mass spectrometry. The modification gave a mass shift of + 70 Da and peptide mapping analysis demonstrated that it located to the protein N-terminus and lysine side chains. The chemical composition of C4H6O was found to be the best candidate by peptide fragmentation using tandem mass spectrometry. The modification likely contains a carbonyl group, since the mass of the modification increased by 2 Da by reduction with borane pyridine complex and it reacted with 2,4-dinitrophenylhydrazine. On the basis of chemical and tandem mass spectrometry fragmentation behavior, the modification could be attributed to crotonaldehyde, a reactive compound formed during lipid peroxidation. A low recorded oxygen pressure in the reactor during protein expression could be linked to the formation of this compound. This study shows the importance of maintaining full control over all reaction parameters during recombinant protein production.

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