1 of 128

Defiance is the Secret of Life�Energy for Fun and Profit

MCB137/237 – Spring 2025

2 of 128

Jacques Monod and the Second Secret of Life

3 of 128

Monod’s Second Secret of Life: Allostery

4 of 128

… But If Allostery Is the Second Secret of Life, What Is the First one?

5 of 128

… But There Are Also Other Contenders

6 of 128

7 of 128

8 of 128

Fidelity in biological polymerization: Key question, are we surprised?

9 of 128

The Insufficiency of Equilibrium Molecular Recognition

10 of 128

What Is the Energy of DNA Pairing?�Intuition from PCR Primer Design

11 of 128

A Toy Model of Translation

12 of 128

The Kinetic Proofreading Idea: Energy to Fuel Error Correction

13 of 128

Translation Does Indeed Require Energy

14 of 128

Life Is Defiance�What Can You Do Only When You Spend Energy?

15 of 128

Life Is Defiance�What Can You Do Only When You Spend Energy?

16 of 128

Viruses Are Constantly Evolving�Tracking Influenza

17 of 128

Tracking the Spread of Coronavirus

18 of 128

Tracking the Spread of Coronavirus

19 of 128

Influenza Evolves Faster Than SARS-CoV-2

20 of 128

Viral Genome Lengths

Mahmoudabadi and Phillips (2018)

21 of 128

A Compromise Between Genome Length and Mutation Rate

Holmes, PNAS (2010)

22 of 128

ssRNA+ Viral Genome Lengths

Gorbalenya et al. (2006)

23 of 128

SARS-CoV-2 +ssRNA Genome

24 of 128

Exonuclease Is Present Only in Viruses With Genomes > 20 kb

25 of 128

Exonuclease Provides Proofreading Activity To SARS-CoV

Eckerle et al., PLoS Pathogen (2010)

26 of 128

The Unreasonable Effectiveness of Statistical Mechanics Models of Bacterial Decision Making

Garcia and Phillips, PNAS (2011);

Brewster et al., Cell (2014); Weinert et al., PRL (2014)

27 of 128

Statistical Mechanics Assumes�No Energy Consumption

28 of 128

Life Is Defiance�What Can You Do Only When You Spend Energy?

29 of 128

The Nature of Life is Confusing:�The Example of Diffusion

Jean Baptiste Perrin, 1909

Robert Brown, 1828

30 of 128

Humpty Dumpty and the Cost of Irreversibility

Humpty Dumpty sat on a wall,�Humpty Dumpty had a great fall.�All the king's horses and all the king's men�Couldn't put Humpty together again

31 of 128

Can We Run the Movie of Life Backwards?

32 of 128

The Birth of Maxwell’s Demon

But if we conceive a being whose faculties are so sharpened that he can follow every molecule in its course, such a being, whose attributes are still as essentially finite as our own, would be able to do what is at present impossible to us. For we have seen that the molecules in a vessel full of air at uniform temperature are moving with velocities by no means uniform, though the mean velocity of any great number of them, arbitrarily selected, is almost exactly uniform. Now let us suppose that such a vessel is divided into two portions, A and B, by a division in which there is a small hole, and that a being, who can see the individual molecules, opens and closes this hole, so as to allow only the swifter molecules to pass from A to B, and only the slower ones to pass from B to A. He will thus, without expenditure of work, raise the temperature of B and lower that of A, in contradiction to the second law of thermodynamics.

From a letter from Maxwell to Tait

33 of 128

Running the Movie of Life Backwards… …If You Are Willing to Pay!

34 of 128

A Huge Amounts of Energy Stored Around Us

35 of 128

Demonic Biology

36 of 128

Our Intuition About Spatial Scales in Biology�(22 orders of magnitude)

37 of 128

Our Intuition About Temporal Scales in Biology�(29 orders of magnitude)

38 of 128

What’s Our Intuition About Energy Scales in Biology?�(43 orders of magnitude!)

39 of 128

Estimate: How Much Energy from Sunlight Hits the Earth in a Day?

40 of 128

41 of 128

What Is the Daily Energy Consumption of a Human?

42 of 128

What Is the Daily Energy Consumption of a Human?

43 of 128

What Is the Daily Energy Consumption of the Human Poulation?

44 of 128

How Much Energy to Build an E. coli cell?

45 of 128

46 of 128

Energy to Build a Protein

47 of 128

48 of 128

49 of 128

50 of 128

Concentrations and Absolute Numbers in Your Favorite Cell Type

51 of 128

52 of 128

53 of 128

Deterministic Versus Thermal Energies�Using Dimensionless Numbers to Get a Sense of Scale

54 of 128

Energy Can Be Converted Between Multiple Forms

55 of 128

Some Examples of Energy Conversion

56 of 128

Human Scheme for Energy Generation

57 of 128

Plant Scheme for Energy Generation

58 of 128

A Hierarchy of Biological Batteries

59 of 128

Molecular Batteries Powering the Cell

60 of 128

Where the Energy Comes From Is None of Your Business!

61 of 128

A Hierarchy of Biological Batteries

62 of 128

63 of 128

64 of 128

Stouthamer’s Analysis of ATP Usage in Bacteria

65 of 128

Energy Requirements Scale With Cellular Volume

Lynch and Marinov (2015)

66 of 128

Synthesizing ATP

67 of 128

ATP Synthase

68 of 128

Rotating at the speed of a jet engine

69 of 128

Synthesizing ATP: Your turn

  • How many ATP synthases do I need to keep up with the energy requirements of the cell?
  • Can I fit all of those molecules on the cell membrane?

70 of 128

Number of ATP Synthases in the Cell

71 of 128

Competition Between Surface Area and Volume in Cellular Energetics

72 of 128

Competition between ATP Consumption and Production Versus Cell Size

73 of 128

An Hypothesis: Mitochondria Circumvent the Competition Between Surface Area and Volume

74 of 128

75 of 128

76 of 128

Concentrations and Absolute Numbers in Your Favorite Cell Type

77 of 128

78 of 128

Musings on Entropy and Life

``The entropy of the whole universe must be increasing. It follows that each of us, as a living organism that locally and temporarily decreases entropy, must produce somewhere in the world around us an increase in entropy. As we metabolize food, for example, we give off heat and increase random molecular motion around us. In a sense, we buy our lives through the entropic death of the universe.'' - Mathews & van Holde (1990), Biochemistry (Prentice Hall).

79 of 128

Defying Diffusion

80 of 128

Denomic Diffusion

81 of 128

The Demon Has to Fight Against Membrane Permeability

82 of 128

Harvesting the Energy Paid by the Demon�The Proton Motive Force

83 of 128

ATP Synthase

84 of 128

Measuring ATP Synthase Rotation

85 of 128

A Simple Model of Membrane Potential and Proton Transport

86 of 128

The ATP Synthase Converts Energy Stored in the Proton Gradient Into ATP

87 of 128

Harnessing Concentration Gradients to Do Work

88 of 128

LacY from the lac Operon Pumps Lactose Into the Cell

89 of 128

LacY Harnesses the H+ Gradient to Pump Lactose In

90 of 128

Symporters Act As AND Gates

91 of 128

Symporters Harness Ion Gradients to Facilitate Molecular Transport

Nsugar = 3

Nion = 8

Nsugar = 9

Nion = 4

Nsugar = 2

Nion = 7

Nsugar = 10

Nion = 5

92 of 128

The exchange made in the cellular economy

93 of 128

A Hierarchy of Biological Batteries

94 of 128

Energy in ATP Hydrolysis

95 of 128

Where Does the Energy in ATP Hydrolysis Come From?

96 of 128

Searching for Non-Equilibrium Signatures in Cellular Decision Making

97 of 128

A Closer Look at Translation Error Rates

98 of 128

The Wider Problem of Molecular Recognition

99 of 128

HP Model of Proteins: A Toy Model to Explore Fidelity

100 of 128

Molecular Recognition in Translation

101 of 128

A Simple Model of Amino Acid Incorporation

102 of 128

Energetics of Codon Recognition

  • 1 H bond is worth about 2 kBT

103 of 128

Molecular Recognition

104 of 128

But Error Rates Are Much Lower Than Predicted by Thermodynamics

105 of 128

Need a Maxwell Demon That Defies Thermodynamics

Rob Phillips

106 of 128

The Kinetic Proofreading Idea

107 of 128

Translation Consumes Energy

108 of 128

A Simplified Model of Kinetic Proofreading

109 of 128

Prominence of One-Way Arrows in Our Cartoon Models of Biology

110 of 128

Biological Assemblies Call for One-Way Arrows

111 of 128

Energy Expenditure Makes It Possible to Encode for Temporal Ordering of Events

112 of 128

A Simplified Model of Kinetic Proofreading

113 of 128

Allostery�Relating Hydrolysis to Mechanical Work

114 of 128

The Demon in the Piston

115 of 128

The Allosteric Transition Gives One More Opportunity for Ligand Discrimination

116 of 128

Waiting Time Distributions of Correct and Incorrect Ligand Unbinding

117 of 128

Revealing Translational Dynamics Using Stochastic Simulations

118 of 128

Confirming Our Prediction About the Fraction of Errors in Translation

119 of 128

Stochastic Simulations�One of the Three Protocols for Solving Biological Dynamics

120 of 128

Trajectories and Probabilities of the Constitutive Promoter

121 of 128

The Stochastic Simulation Protocol

122 of 128

Stochastic Simulation of the Constitutive Promoter

123 of 128

A Hierarchy of Biological Batteries

124 of 128

Life Is Defiance�What Can You Do Only When You Spend Energy?

125 of 128

Prominence of One-Way Arrows in Our Cartoon Models of Biology

126 of 128

Biological Assemblies Call for One-Way Arrows

127 of 128

Energy Expenditure Makes It Possible to Encode for Temporal Ordering of Events

128 of 128

SLIDE LEFT BLANK INTENTIONALLY