scholarly journals Rapid generation of incremental truncation libraries for protein engineering using alpha-phosphothioate nucleotides

2001 ◽  
Vol 29 (4) ◽  
pp. 16e-16 ◽  
Author(s):  
S. Lutz
Keyword(s):  
Protein Engineering ◽  
Incremental Truncation ◽  
Rapid Generation

scholarly journals Combinatorial protein engineering by incremental truncation

1999 ◽  
Vol 96 (7) ◽  
pp. 3562-3567 ◽  
Author(s):  
M. Ostermeier ◽  
A. E. Nixon ◽  
J. H. Shim ◽  
S. J. Benkovic
Keyword(s):  
Protein Engineering ◽  
Combinatorial Protein Engineering ◽  
Incremental Truncation

Insights into the biochemical and functional characterization of sortase E transpeptidase of Corynebacterium glutamicum

2019 ◽  
Vol 476 (24) ◽  
pp. 3835-3847 ◽  
Author(s):  
Aliyath Susmitha ◽  
Kesavan Madhavan Nampoothiri ◽  
Harsha Bajaj
Keyword(s):  
Protein Engineering ◽  
Cell Attachment ◽  
Functional Characterization ◽  
Protein Structures ◽  
Structural Similarity ◽  
Surface Proteins ◽  
Sortase A ◽  
Peptide Cleavage ◽  
New Findings

Most Gram-positive bacteria contain a membrane-bound transpeptidase known as sortase which covalently incorporates the surface proteins on to the cell wall. The sortase-displayed protein structures are involved in cell attachment, nutrient uptake and aerial hyphae formation. Among the six classes of sortase (A–F), sortase A of S. aureus is the well-characterized housekeeping enzyme considered as an ideal drug target and a valuable biochemical reagent for protein engineering. Similar to SrtA, class E sortase in GC rich bacteria plays a housekeeping role which is not studied extensively. However, C. glutamicum ATCC 13032, an industrially important organism known for amino acid production, carries a single putative sortase (NCgl2838) gene but neither in vitro peptide cleavage activity nor biochemical characterizations have been investigated. Here, we identified that the gene is having a sortase activity and analyzed its structural similarity with Cd-SrtF. The purified enzyme showed a greater affinity toward LAXTG substrate with a calculated KM of 12 ± 1 µM, one of the highest affinities reported for this class of enzyme. Moreover, site-directed mutation studies were carried to ascertain the structure functional relationship of Cg-SrtE and all these are new findings which will enable us to perceive exciting protein engineering applications with this class of enzyme from a non-pathogenic microbe.

Early Platelet-Collagen Interactions in Whole Blood and Their Modifications by Aspirin and Dipyridamole Evaluated by a New Method (Basic Wave)

1985 ◽  
Vol 54 (04) ◽  
pp. 799-803 ◽  
Author(s):  
José Luís Pérez-Requejo ◽  
Justo Aznar ◽  
M Teresa Santos ◽  
Juana Vallés
Keyword(s):  
Whole Blood ◽  
Ex Vivo ◽  
Platelet Rich Plasma ◽  
White Blood Cells ◽  
Reversible Aggregation ◽  
Inhibitory Compounds ◽  
Rapid Generation ◽  
Stimulation Of ◽  
Collagen Stimulation

SummaryIt is shown that the supernatant of unstirred whole blood at 37° C, stimulated by 1 μg/ml of collagen for 10 sec, produces a rapid generation of pro and antiaggregatory compounds with a final proaggregatory activity which can be detected for more than 60 min on a platelet rich plasma (PRP) by turbidometric aggregometry. A reversible aggregation wave that we have called BASIC wave (for Blood Aggregation Stimulatory and Inhibitory Compounds) is recorded. The collagen stimulation of unstirred PRP produces a similar but smaller BASIC wave. BASIC’s intensity increases if erythrocytes are added to PRP but decreases if white blood cells are added instead. Aspirin abolishes “ex vivo” the ability of whole blood and PRP to generate BASIC waves and dipyridamole “in vitro” significantly reduces BASIC’s intensity in whole blood in every tested sample, but shows little effect in PRP.

Protein Engineering of Penicillin Acylase

Acta Naturae ◽  
2010 ◽  
Vol 2 (3) ◽  
pp. 47-61 ◽  
Author(s):  
V I Tishkov ◽  
S S Savin ◽  
A S Yasnaya
Keyword(s):  
Protein Engineering ◽  
Penicillin Acylase

Rapid Failure Analysis of Low-Yielding Electrical Test Structures Using E-beam Physical and Voltage Contrast Inspection

2013 ◽  
Author(s):  
Oliver D. Patterson ◽  
Deborah A. Ryan ◽  
Xiaohu Tang ◽  
Shuen Cheng Lei
Keyword(s):  
Failure Analysis ◽  
Defect Detection ◽  
High Volume ◽  
The Other ◽  
Defect Inspection ◽  
Electrical Test ◽  
Detection Mode ◽  
Rapid Generation ◽  
Voltage Contrast ◽  
Additional Support

Abstract In-line E-beam inspection may be used for rapid generation of failure analysis (FA) results for low yielding test structures. This approach provides a number of advantages: 1) It is much earlier than traditional FA, 2) de-processing isn’t required, and 3) a high volume of sites can be processed with the additional support of an in-line FIB. Both physical defect detection and voltage contrast inspection modes are useful for this application. Voltage contrast mode is necessary for isolation of buried defects and is the preferred approach for opens, because it is faster. Physical defect detection mode is generally necessary to locate shorts. The considerations in applying these inspection modes for rapid failure analysis are discussed in the context of two examples: one that lends itself to physical defect inspection and the other, more appropriately addressed with voltage contrast inspection.

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