Review of: \u3cem\u3eCarpenter Under Construction: The Story of Don Plank\u3c/em\u3e—Diane Freed

Carpenter Under Construction is the story of Don Plank’s life, written by his youngest daughter. It briefly touches on his early life and moves through major life events. About half of the book covers the busy later years in life when he and his second wife moved from mission to mission in various support roles. Don was a good carpenter, but Freed puts more emphasis on what God built Don to be rather than on what Don himself built during his long life. [First paragraph.

A tale of three companies

No Abstracts

Cell-free synthesis of membrane bound polypeptides

Methods are provided for the utilization of bacterial cell-free extracts in the synthesis of high yields of membrane-associated polypeptides

Cell-free production of active E. coli thioredoxin reductase and glutathione reductase

AbstractEscherichia coli thioredoxin reductase (TR) and glutathione reductase (GR) are dimeric proteins that require a flavin adenine dinucleotide (FAD) cofactor for activity. A cell-free protein synthesis (CFPS) reaction supplemented with FAD was used to produce TR at 760 μg/ml with 89% of the protein being soluble. GR accumulated to 521 μg/ml in a cell-free reaction with 71% solubility. The TR produced was fully active with a specific activity of 1390 min−1. The GR had a specific activity of 139 U/mg, which is significantly more active than reported for GR purified from cells. The specific activity for both TR and GR decreased without FAD supplementation. This research demonstrates that CFPS can be used to produce enzymes that are multimeric and require a cofactor

Tyrosine, Cysteine, and S-Adenosyl Methionine Stimulate In Vitro [FeFe] Hydrogenase Activation

Background: [FeFe] hydrogenases are metalloenzymes involved in the anaerobic metabolism of H2. These proteins are distinguished by an active site cofactor known as the H-cluster. This unique [6Fe–6S] complex contains multiple non-protein moieties and requires several maturation enzymes for its assembly. The pathways and biochemical precursors for H-cluster biosynthesis have yet to be elucidated. Principal Findings: We report an in vitro maturation system in which, for the first time, chemical additives enhance [FeFe] hydrogenase activation, thus signifying in situ H-cluster biosynthesis. The maturation system is comprised of purified hydrogenase apoprotein; a dialyzed Escherichia coli cell lysate containing heterologous HydE, HydF, and HydG maturases; and exogenous small molecules. Following anaerobic incubation of the Chlamydomonas reinhardtii HydA1 apohydrogenase with S-adenosyl methionine (SAM), cysteine, tyrosine, iron, sulfide, and the non-purified maturases, hydrogenase activity increased 5-fold relative to incubations without the exogenous substrates. No conditions were identified in which addition of guanosine triphosphate (GTP) improved hydrogenase maturation. Significance: The in vitro system allows for direct investigation of [FeFe] hydrogenase activation. This work also provides a foundation for studying the biosynthetic mechanisms of H-cluster biosynthesis using solely purified enzymes and chemical additives

Contextual batting and bowling in limited overs cricket

Cricket is a sport for which many batting and bowling statistics have been proposed. However, a feature of cricket is that the level of aggressiveness adopted by batsmen is dependent on match circumstances. It is therefore relevant to consider these circumstances when evaluating batting and bowling performances. This paper considers batting performance in the second innings of limited overs cricket when a target has been set. The runs required, the number of overs completed and the wickets taken are relevant in assessing the batting performance. We produce a visualization for second innings batting which describes how a batsman performs under different circumstances. The visualization is then reduced to a single statistic “clutch batting” which can be used to compare batsmen. An analogous approach is then provided for bowlers based on the symmetry between batting and bowling, and we define the statistic “clutch bowling”