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From Biochemistry to Physiology: The Calorimetry Connection

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Abstract

This article provides guidelines for selecting optimal calorimetric instrumentation for applications in biochemistry and biophysics. Applications include determining thermodynamics of interactions in non-covalently bonded structures, and determining function through measurements of enzyme kinetics and metabolic rates. Specific examples illustrating current capabilities and methods in biological calorimetry are provided. Commercially available calorimeters are categorized by application and by instrument characteristics (isothermal or temperature-scanning, reaction vessel volume, heat rate detection limit, fixed or removable reaction vessels, etc.). Advantages and limitations of commercially available calorimeters are listed for each application in biochemistry, biophysics, and physiology.

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Acknowledgments

LDH thanks BYU for continuing support. We thank Kevin Peine for growing the pumpkins and collecting the metabolic data on leaf tissue.

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Authors and Affiliations

  1. Calorimetry Sciences Corp., 890 W 410 N, Suite A, Lindon, UT, 84042, USA

    Lee D. Hansen, Donald J. Russell & Christin T. Choma

  2. Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 84602, USA

    Lee D. Hansen

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Correspondence to Donald J. Russell.

Appendix

Appendix

Measurement of Heat

There are only three ways of measuring heat; these are known as temperature rise, heat conduction, and power compensation methods [63]. In the temperature rise method, the temperature change caused by a process is multiplied by the apparent heat capacity of the calorimetric system to obtain the heat effect. The reaction vessel is typically a glass Dewar that may have a time constant as low as a fraction of a second. Although this method is inherently the fastest method for heat measurement, and has been extensively applied to binding reactions in multi-reaction systems [21] and for measurement of kinetics, it is limited to short measurement time periods (< 2 h) and relatively fast reactions because of the need to correct for heat exchange with the environment. Also, because use of very small volumes complicates heat exchange corrections [64], the temperature rise method generally requires more material for experiments than the other methods. In the heat conduction method, heat rate is calculated from the measured temperature difference across a fixed thermal conductivity path, i.e., dQ/dt = kΔT. The ΔT is commonly measured as a voltage output from a Seebeck device or thermopile between the sample chamber and a metal block at a reference temperature, which is either held constant or scanned according to a preset program. In the power compensation method, the power from an electrical heater required to maintain the sample at a reference temperature is measured. Again the reference temperature may be held constant or scanned. Although power compensation is generally a faster method than heat conduction if the methods are compared at similar detection limits and sample size, this choice is probably of little consequence in selecting a calorimeter for the applications discussed in this article.

The surroundings of a calorimeter vessel can be isoperibol (literally: constant surroundings) or adiabatic (the temperature of the surroundings is controlled to be equal to the temperature of the reaction vessel). Isoperibol systems are simpler to build and maintain than adiabatic systems, and are capable of producing equally good data.

Nomenclature and Units

In this article the term “heat rate” is used for dQ/dt instead of “thermal power” because of possible confusion caused by the latter term. The heat rate should be expressed in the SI system of units, for example, in microwatts or microjoules per second, and not as microcalories per second. The calorie, 4.184 J, is not an acceptable SI unit, and many journals now require data to be reported in SI units.

Another source of confusion is use of the terms “oxycaloric equivalent”, “calorespirometric ratio”, and “Thornton’s rule” or “Thornton’s constant”. The oxycaloric equivalent is the amount of heat produced per mole of oxygen consumed by an aerobic, living system in a steady state condition, and is typically near 455 kJ per mole of O2. Thornton’s constant is derived from heats of combustion of organic compounds [65]. Since it is not truly a constant, i.e., typically this value is 455 ± 15 kJ per mole of O2, it is more properly referred to as a rule rather than a constant. The exact value depends on the substrate being oxidized and on the state of the reactants and products.

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Hansen, L.D., Russell, D.J. & Choma, C.T. From Biochemistry to Physiology: The Calorimetry Connection. Cell Biochem Biophys 49, 125–140 (2007). https://doi.org/10.1007/s12013-007-0049-y

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