Heads of Laboratories
Investigator, Howard Hughes Medical Institute
John D. Rockefeller Jr. Professor
Laboratory of Molecular Neurobiology and Biophysics
Ion channels catalyze the diffusion of inorganic ions down their electrochemical gradients across cell membranes. Because the ionic movements are passive, ion channels would seem to be extraordinarily simple physical systems, yet they are responsible for electrical signaling in living cells. Among their many functions, ion channels control the pace of the heart, regulate the secretion of hormones into the bloodstream, and generate the electrical impulses underlying information transfer in the nervous system. Dr. MacKinnon's research is aimed at understanding the physical and chemical principles underlying ion channel function.
Chloride channels and transporters mediate the conduction of chloride ions across cell membranes. To understand the chemistry of anion selectivity, Dr. MacKinnon and his colleagues determined the atomic structures of two prokaryotic and one eukaryotic member of the CLC chloride channel family. These anion-selective transport proteins are built on a completely different architectural plan than that of potassium channels, but they nevertheless use many of the same basic physical principles to catalyze selective ionic movement across the membrane. These principles of selective ion movement include oriented alpha-helical-end charges to stabilize ions inside the membrane and short selectivity filters containing multiple ions in a queue to mediate conduction. These structures have also led to an atomic model that explains how CLC transporters couple the counter transport of two chloride ions per single proton across the cell membrane.
Ion Channel Gating
In humans, 78 unique potassium channel genes underlie an impressive level of functional diversity. The different potassium channels have evolved the ability to open — by a process called gating — in response to different forces in their environment. Certain potassium channels are gated by the binding of calcium ions, specific membrane lipids, or G protein subunits. Others respond to mechanical perturbations of the membrane or changes in voltage across the membrane. Thus, the potassium channels control a cell’s electrical state through many different avenues.
To understand the atomic basis of potassium channel gating, the MacKinnon lab has analyzed the function and determined the structures of several different members of the potassium channel family. MthK is a prokaryotic calcium-gated potassium channel, and the BK channel is a eukaryotic cousin of MthK that regulates vascular and airway smooth muscle tone in mammals. Through studies of these channels they have begun to understand how the free energy of calcium binding is converted to mechanical work to open the pore.
Inward rectifier potassium (Kir) channels control the resting membrane voltage in many neuronal and muscle cells. The lab has recently determined atomic structures to understand how the signaling lipid PIP2 regulates gating in Kir2 (a lipid gated channel) and how G proteins regulate gating in Kir3 (a G protein gated channel). In Kir2, when PIP2 binds to the channel it causes a large cytoplasmic domain to move 6 Å to a position that opens the pore. In essence, PIP2 serves as molecular Velcro to tether two components of the channel together, a requirement to open the conduction pathway. In Kir3, large G protein subunits (b and g subunits) bind to the channel at the intracellular interface where the channel meets the cell’s interior. When the G protein subunits bind they cause a twisting of the a helices that form the pore’s gate, and begin to open it. The structure of Kir3 with the G protein subunits bound complete a story that began in 1921, when Otto Loewi discovered that vagus nerve stimulation slows the heart rate. In Dr. Loewi’s experiments, the neurotransmitter acetylcholine was released from the nerve and activated a G protein coupled receptor in the heart pacemaker cells. The G proteins bound to Kir3 and opened it, by a mechanism revealed now in atomic detail, to hyperpolarize the pacemaker cell membrane and slow the heart rate.
Probably the most difficult and in many ways fascinating form of gating the group has studied is called voltage-dependent gating. This gating allows "excitable" cells to produce electrical impulses called action potentials: these underlie muscle contraction and nervous system function. Dr. MacKinnon's group has determined structures of both archaebacterial and eukaryotic voltage-dependent potassium (Kv) channels. In 2005, they published the structure of the Kv1.2 channel from rat, which was the first recombinant eukaryotic membrane protein structure ever solved by x-ray crystallography. Since, they have solved numerous structures to deduce mechanism. Based on these findings they have posited several fundamentally new ideas: (1) voltage-sensor domains are loosely attached to the pore at its perimeter, (2) they contain a helix-turn-helix element that they named the voltage-sensor paddle, (3) the voltage-sensor moves in intimate contact with the membrane, carrying with it charges within the membrane electric field, and (4) the voltage sensor is attached to levers that mechanically open and close the pore. These structures are allowing researchers to begin to understand how nature, through its ion channel voltage sensors, has evolved the analog of a ‘field effect transistor’ to produce action potentials, which have permitted rapid information transfer in living systems. It is fascinating to realize that voltage-dependent gating, which at a cellular level would seem to be the most complex form of gating, at a molecular level might be the simplest: the voltage across the membrane sets up an electric field that pulls and pushes a charged voltage sensor that is attached to a lever that opens and closes the pore.
Most recently Dr. MacKinnon's group has begun to study a different form of gating called mechanical gating. They would like to understand how mechanical forces elicit electrical signals. This form of gating underlies sensory detection as in touch and hearing. It also underlies basic physiological processes such as cell volume regulation and blood pressure control. Dr. MacKinnon and others have observed significant mechanosensitivity in the gating of certain potassium and calcium channels. Now they seek to understand whether mechanosensitivity is physiologically relevant in potassium and calcium channels. They have also determined atomic structures of the human TRAAK channel, which is strongly mechanosensitive, to understand how it transduces mechanical forces, and why the central nervous system expresses a mechanosensitive potassium channel.
The family of potassium channels provides an incredible example of molecular diversity. In an attempt to understand these molecules one never ceases to be amazed by life’s simple solutions to what at first appear to be complicated tasks.
B.A. in biochemistry, 1978
Tufts University School of Medicine
Residency in internal medicine, 1982–1985
Beth Israel Hospital, Harvard Medical School
Harvard University, 1985–1986
Brandeis University, 1986–1989
Assistant Professor, 1989–1992
Associate Professor, 1992–1995
Harvard Medical School
The Rockefeller University
Howard Hughes Medical Institute.
Albert Lasker Basic Medical Research Award, 1999
Lewis S. Rosenstiel Award, 2000
Canada Gairdner International Award, 2001
Perl-UNC Neuroscience Prize, 2001
Nobel Prize in Chemistry, 2003
Louisa Gross Horwitz Prize, 2003
National Academy of Sciences
American Philosophical Society
Hite, R.K. et al. Cryo-electron microscopy structure of the Slo2.2 Na⁺-activated K⁺ channel. Nature 527, 198–203 (2015).
Brohawn, S.G., et al. Physical mechanism for gating and mechanosensitivity of the human TRAAK K⁺ channel. Nature 516, 126–130 (2014).
Wang, W., et al. Quantitative analysis of mammalian GIRK2 channel regulation by G proteins, the signaling lipid PIP2 and Na⁺ in a
reconstituted system. eLife 3, e03671 (2014).
Whorton, M.R. and MacKinnon, R. X-ray structure of the mammalian GIRK2- G-protein complex. Nature 498, 190–197 (2013).
Brohawn, S.G., et al. Crystal structure of the human K2P TRAAK, a lipid- and mechano-sensitive K⁺ ion channel. Science 335,
Dr. MacKinnon is a faculty member in the David Rockefeller Graduate Program, the Tri-Institutional M.D.-Ph.D. Program, and the Tri-Institutional Ph.D. Program in Chemical Biology.