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Chapter 19 Oxidative Phosphorylation
Living Graph:
Free-Energy Change for Transport of an Ion
David L. Nelson
Professor Emeritus of Biochemistry
University of Wisconsin–Madison
Michael M. Cox
Professor of Biochemistry
University of Wisconsin–Madison
Vice President, STEM: Ben Roberts
Senior Acquisitions Editor: Lauren Schultz
Senior Developmental Editor: Susan Moran
Assistant Editor: Shannon Moloney
Marketing Manager: Maureen Rachford
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Director of Media and Assessment: Amanda Nietzel
Media Editor: Lori Stover
Director of Content (Sapling Learning): Clairissa Simmons
Lead Content Developer, Biochemistry (Sapling Learning): Richard Widstrom
Content Development Manager for Chemistry (Sapling Learning): Stacy Benson
Visual Development Editor (Media): Emiko Paul
Director, Content Management Enhancement: Tracey Kuehn
Managing Editor: Lisa Kinne
Senior Project Editor: Liz Geller
Copyeditor: Linda Strange
Photo Editor: Christine Buese
Photo Researcher: Roger Feldman
Text and Cover Design: Blake Logan
Illustration Coordinator: Janice Donnola
Illustrations: H. Adam Steinberg
Molecular Graphics: H. Adam Steinberg
Production Manager: Susan Wein
Composition: Aptara, Inc.
Printing and Binding: RR Donnelley
Front Cover Image: H. Adam Steinberg and Quade Paul
Back Cover Photo: Yigong Shi
Front cover: An active spliceosome from the yeast Schizosaccharomyces pombe. The
structure, determined by cryo-electron microscopy, captures a molecular moment when the
splicing reaction is nearing completion. It includes the snRNAs U2, U5, and U6, a spliced
intron lariat, and many associated proteins. Structure determined by Yigong Shi and
colleagues, Tsinghua University, Beijing, China (PDB ID 3JB9, C. Yan et al., Science
349:1182, 2015). Back cover: Randomly deposited individual spliceosome particles,
viewed by electron microscopy. The structure on the front cover was obtained by
computationally finding the orientations that are superposable, to reduce the noise and
strengthen the signal—the structure of the spliceosome. Photo courtesy of Yigong Shi.
First printing
International Edition
Macmillan Higher Education
Houndmills, Basingstoke
RG21 6XS, England
www.macmillanhighered.com/international
To Our Teachers
Paul R. Burton
Albert Finholt
William P. Jencks
Eugene P. Kennedy
Homer Knoss
Arthur Kornberg
I. Robert Lehman
Earl K. Nelson
Wesley A. Pearson
David E. Sheppard
Harold B. White
About the Authors
Mike Cox has coordinated a large and active research team at Wisconsin,
investigating the enzymology, topology, and energetics of the
recombinational DNA repair of double-strand breaks in DNA. The work has
focused on the bacterial RecA protein, a wide range of proteins that play
auxiliary roles in recombinational DNA repair, the molecular basis of
extreme resistance to ionizing radiation, directed evolution of new
phenotypes in bacteria, and the applications of all of this work to
biotechnology.
For more than three decades he has taught a survey of biochemistry to
undergraduates and has lectured in graduate courses on DNA structure and
topology, protein-DNA interactions, and the biochemistry of recombination.
More recent projects are the organization of a new course on professional
responsibility for first-year graduate students and establishment of a
systematic program to draw talented biochemistry undergraduates into the
laboratory at an early stage of their college career. He has received awards for
both his teaching and his research, including the Dreyfus Teacher–Scholar
Award, the 1989 Eli Lilly Award in Biological Chemistry, and the 2009
Regents Teaching Excellence Award from the University of Wisconsin. He is
also highly active in national efforts to provide new guidelines for
undergraduate biochemistry education. Cox’s hobbies include turning 18
acres of Wisconsin farmland into an arboretum, wine collecting, and assisting
in the design of laboratory buildings.
A Note on the Nature of
Science
I
n this twenty-first century, a typical science education often leaves the
philosophical underpinnings of science unstated, or relies on
oversimplified definitions. As you contemplate a career in science, it may
be useful to consider once again the terms science, scientist, and scientific
method.
Science is both a way of thinking about the natural world and the sum of
the information and theory that result from such thinking. The power and
success of science flow directly from its reliance on ideas that can be tested:
information on natural phenomena that can be observed, measured, and
reproduced and theories that have predictive value. The progress of science
rests on a foundational assumption that is often unstated but crucial to the
enterprise: that the laws governing forces and phenomena existing in the
universe are not subject to change. The Nobel laureate Jacques Monod
referred to this underlying assumption as the “postulate of objectivity.” The
natural world can therefore be understood by applying a process of inquiry—
the scientific method. Science could not succeed in a universe that played
tricks on us. Other than the postulate of objectivity, science makes no
inviolate assumptions about the natural world. A useful scientific idea is one
that (1) has been or can be reproducibly substantiated, (2) can be used to
accurately predict new phenomena, and (3) focuses on the natural world or
universe.
Scientific ideas take many forms. The terms that scientists use to describe
these forms have meanings quite different from those applied by
nonscientists. A hypothesis is an idea or assumption that provides a
reasonable and testable explanation for one or more observations, but it may
lack extensive experimental substantiation. A scientific theory is much more
than a hunch. It is an idea that has been substantiated to some extent and
provides an explanation for a body of experimental observations. A theory
can be tested and built upon and is thus a basis for further advance and
innovation. When a scientific theory has been repeatedly tested and validated
on many fronts, it can be accepted as a fact.
In one important sense, what constitutes science or a scientific idea is
defined by whether or not it is published in the scientific literature after peer
review by other working scientists. As of late 2014, about 34,500 peer-
reviewed scientific journals worldwide were publishing some 2.5 million
articles each year, a continuing rich harvest of information that is the
birthright of every human being.
Scientists are individuals who rigorously apply the scientific method to
understand the natural world. Merely having an advanced degree in a
scientific discipline does not make one a scientist, nor does the lack of such a
degree prevent one from making important scientific contributions. A
scientist must be willing to challenge any idea when new findings demand it.
The ideas that a scientist accepts must be based on measurable, reproducible
observations, and the scientist must report these observations with complete
honesty.
The scientific method is a collection of paths, all of which may lead to
scientific discovery. In the hypothesis and experiment path, a scientist poses a
hypothesis, then subjects it to experimental test. Many of the processes that
biochemists work with every day were discovered in this manner. The DNA
structure elucidated by James Watson and Francis Crick led to the hypothesis
that base pairing is the basis for information transfer in polynucleotide
synthesis. This hypothesis helped inspire the discovery of DNA and RNA
polymerases.
Watson and Crick produced their DNA structure through a process of
model building and calculation. No actual experiments were involved,
although the model building and calculations used data collected by other
scientists. Many adventurous scientists have applied the process of
exploration and observation as a path to discovery. Historical voyages of
discovery (Charles Darwin’s 1831 voyage on H.M.S. Beagle among them)
helped to map the planet, catalog its living occupants, and change the way we
view the world. Modern scientists follow a similar path when they explore
the ocean depths or launch probes to other planets. An analog of hypothesis
and experiment is hypothesis and deduction. Crick reasoned that there must
be an adaptor molecule that facilitated translation of the information in
messenger RNA into protein. This adaptor hypothesis led to the discovery of
transfer RNA by Mahlon Hoagland and Paul Zamecnik.
Not all paths to discovery involve planning. Serendipity often plays a
role. The discovery of penicillin by Alexander Fleming in 1928 and of RNA
catalysts by Thomas Cech in the early 1980s were both chance discoveries,
albeit by scientists well prepared to exploit them. Inspiration can also lead to
important advances. The polymerase chain reaction (PCR), now a central part
of biotechnology, was developed by Kary Mullis after a flash of inspiration
during a road trip in northern California in 1983.
These many paths to scientific discovery can seem quite different, but
they have some important things in common. They are focused on the natural
world. They rely on reproducible observation and/or experiment. All of the
ideas, insights, and experimental facts that arise from these endeavors can be
tested and reproduced by scientists anywhere in the world. All can be used by
other scientists to build new hypotheses and make new discoveries. All lead
to information that is properly included in the realm of science.
Understanding our universe requires hard work. At the same time, no human
endeavor is more exciting and potentially rewarding than trying, with
occasional success, to understand some part of the natural world.
Preface
W
ith the advent of increasingly robust technologies that provide
cellular and organismal views of molecular processes, progress in
biochemistry continues apace, providing both new wonders and new
challenges. The image on our cover depicts an active spliceosome, one of the
largest molecular machines in a eukaryotic cell, and one that is only now
yielding to modern structural analysis. It is an example of our current
understanding of life at the level of molecular structure. The image is a
snapshot from a highly complex set of reactions, in better focus than ever
before. But in the cell, this is only one of many steps linked spatially and
temporally to many other complex processes that remain to be unraveled and
eventually described in future editions. Our goal in this seventh edition of
Lehninger Principles of Biochemistry, as always, is to strike a balance: to
include new and exciting research findings without making the book
overwhelming for students. The primary criterion for inclusion of an advance
is that the new finding helps to illustrate an important principle of
biochemistry.
With every revision of this textbook, we have striven to maintain the
qualities that made the original Lehninger text a classic: clear writing, careful
explanations of difficult concepts, and insightful communication to students
of the ways in which biochemistry is understood and practiced today. We
have coauthored this text and taught introductory biochemistry together for
three decades. Our thousands of students at the University of Wisconsin–
Madison over those years have been an endless source of ideas on how to
present biochemistry more clearly; they have enlightened and inspired us. We
hope that this seventh edition of Lehninger will, in turn, enlighten current
students of biochemistry everywhere, and inspire all of them to love
biochemistry as we do.
■ Online data resources such as NCBI, PDB, SCOP2, KEGG, and BLAST,
mentioned in the text, are listed in the back endpapers for easy reference.
CRISPR/Cas9 structure
Problem-Solving Tools
■ In-text Worked Examples help students improve their quantitative
problem-solving skills, taking them through some of the most difficult
equations.
■ More than 600 end-of-chapter problems give students further
opportunity to practice what they have learned.
■ Data Analysis Problems (one at the end of each chapter), contributed by
Brian White of the University of Massachusetts Boston, encourage students
to synthesize what they have learned and apply their knowledge to
interpretation of data from the research literature.
Key Conventions
Many of the conventions that are so necessary for understanding each
biochemical topic and the biochemical literature are broken out of the text
and highlighted. These Key Conventions include clear statements of many
assumptions and conventions that students are often expected to assimilate
without being told (for example, peptide sequences are written from amino-
to carboxyl-terminal end, left to right; nucleotide sequences are written from
5′ to 3′ end, left to right).
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“gives me the tongue of instruction, so as to know in season when it
is requisite to speak a word;”[1118] not in the way of testimony alone,
but also in the way of question and answer. “And the instruction of
the Lord opens my mouth.”[1119] It is the prerogative of the Gnostic,
then, to know how to make use of speech, and when, and how, and
to whom. And already the apostle, by saying, “After the rudiments of
the world, and not after Christ,”[1120] makes the asseveration that the
Hellenic teaching is elementary, and that of Christ perfect, as we
have already intimated before.
“Now the wild olive is inserted into the fatness of the olive,”[1121]
and is indeed of the same species as the cultivated olives. For the
graft uses as soil the tree in which it is engrafted. Now all the plants
sprouted forth simultaneously in consequence of the divine order.
Wherefore also, though the wild olive be wild, it crowns the Olympic
victors. And the elm teaches the vine to be fruitful, by leading it up to
a height. Now we see that wild trees attract more nutriment, because
they cannot ripen. The wild trees, therefore, have less power of
secretion than those that are cultivated. And the cause of their
wildness is the want of the power of secretion. The engrafted olive
accordingly receives more nutriment from its growing in the wild one;
and it gets accustomed, as it were, to secrete the nutriment,
becoming thus assimilated[1122] to the fatness of the cultivated tree.
So also the philosopher, resembling the wild olive, in having much
that is undigested, on account of his devotion to the search, his
propensity to follow, and his eagerness to seize the fatness of the
truth; if he get besides the divine power, through faith, by being
transplanted into the good and mild knowledge, like the wild olive,
engrafted in the truly fair and merciful Word, he both assimilates the
nutriment that is supplied, and becomes a fair and good olive tree.
For engrafting makes worthless shoots noble, and compels the
barren to be fruitful by the art of culture and by gnostic skill.
writes a poet of note, teaching us that the ancient lyre was seven-
toned. The organs of the senses situated on our face are also seven
—two eyes, two passages of hearing, two nostrils, and the seventh
the mouth.
And that the changes in the periods of life take place by sevens,
the Elegies of Solon teach thus:
“The child, while still an infant, in seven years,
Produces and puts forth its fence of teeth;
And when God seven years more completes,
He shows of puberty’s approach the signs;
And in the third, the beard on growing cheek
With down o’erspreads the bloom of changing skin;
And in the fourth septenniad, at his best
In strength, of manliness he shows the signs;
And in the fifth, of marriage, now mature,
And of posterity, the man bethinks;
Nor does he yet desire vain works to see.
The seventh and eighth septenniads see him now
In mind and speech mature, till fifty years;
And in the ninth he still has vigour left,
But strength and body are for virtue great
Less than of yore; when, seven years more, God brings
To end, then not too soon may he submit to die.”
Again, in diseases the seventh day is that of the crisis; and the
fourteenth, in which nature struggles against the causes of the
diseases. And a myriad such instances are adduced by Hermippus
of Berytus, in his book On the Number Seven, regarding it as holy.
And the blessed David delivers clearly to those who know the mystic
account of seven and eight, praising thus: “Our years were exercised
like a spider. The days of our years in them are seventy years; but if
in strength, eighty years. And that will be to reign.”[1158] That, then,
we may be taught that the world was originated, and not suppose
that God made it in time, prophecy adds: “This is the book of the
generation: also of the things in them, when they were created in the
day that God made heaven and earth.”[1159] For the expression
“when they were created” intimates an indefinite and dateless
production. But the expression “in the day that God made,” that is, in
and by which God made “all things,” and “without which not even
one thing was made,” points out the activity exerted by the Son. As
David says, “This is the day which the Lord hath made; let us be glad
and rejoice in it;”[1160] that is, in consequence of the knowledge
imparted by Him, let us celebrate the divine festival; for the Word
that throws light on things hidden, and by whom each created thing
came into life and being, is called day.
And, in fine, the Decalogue, by the letter Iota,[1161] signifies the
blessed name, presenting Jesus, who is the Word.
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