| Chapter title |
An Improved Kinetic Assay for the Characterization of Metal-Dependent Pectate Lyases
|
|---|---|
| Chapter number | 4 |
| Book title |
Protein-Carbohydrate Interactions
|
| Published in |
Methods in molecular biology, April 2017
|
| DOI | 10.1007/978-1-4939-6899-2_4 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6898-5, 978-1-4939-6899-2
|
| Authors |
Darryl R. Jones, Richard McLean, D. Wade Abbott |
| Editors |
D. Wade Abbott, Alicia Lammerts van Bueren |
| Abstract |
Pectate lyases are a subset of polysaccharide lyases (PLs) that specifically utilize a metal dependent β-elimination mechanism to cleave glyosidic bonds in homogalacturonan (HG; α-D-1,4-galacturonic acid). Most commonly, PLs harness calcium for catalysis; however, some PL families (e.g., PL2 and PL22) display preferences for transitional metals. Deploying alternative metals during β-elimination is correlated with signature coordination pocket chemistry, and is reflective of the evolution, functional specialization, and cellular location of PL activity. Here we describe an optimized method for the analysis of metal-dependent polysaccharide lyases (PLs). We use an endolytic PL2 from Yersinia enterocolitica (YePL2A) as example to demonstrate how altering the catalytic metal within the reaction can modulate PL kinetics. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 4 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 2 | 50% |
| Student > Master | 1 | 25% |
| Unknown | 1 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 2 | 50% |
| Agricultural and Biological Sciences | 1 | 25% |
| Unknown | 1 | 25% |