GroEL Locked in a Closed Conformation by an Interdomain Cross-link Can Bind ATP and Polypeptide but Cannot Process Further Reaction Steps

Murai, Noriyuki; Makino, Yoshihide; Yoshida, Masasuke

doi:10.1074/jbc.271.45.28229

Cited by 49 publications

(51 citation statements)

References 43 publications

(39 reference statements)

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“…The cross-linking of two residues in the structure of a protein that are separated by a large number of residues results in a covalently closed large loop within the primary sequence. Denatured proteins containing such a loop might be expected to exhibit a different mobility during gel electrophoresis; examples have been found where cross-linking induces either a gel mobility increase (17) or a gel mobility decrease (18). Fig.…”

Section: Resultsmentioning

confidence: 99%

The Myosin Relay Helix to Converter Interface Remains Intact throughout the Actomyosin ATPase Cycle

Shih¹,

Spudich

2001

Journal of Biological Chemistry

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Myosins are molecular motors that transduce the chemical energy of ATP hydrolysis to mechanical work in the form of the vectorial translocation of substrate actin filaments. In the swinging lever arm model of actin-myosin motor action, myosin binds to the actin with its globular catalytic domain and then rotates its carboxyl-terminal lever arm domain (reviewed in Ref. 5). The anchoring of the end of the lever arm domain results in the translocation of the catalytic domain and the attached actin filament.

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Section: Resultsmentioning

confidence: 99%

The Myosin Relay Helix to Converter Interface Remains Intact throughout the Actomyosin ATPase Cycle

Shih¹,

Spudich

2001

Journal of Biological Chemistry

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“…Helix M contributes the carboxylate oxygen of Asp398 to the Mg 2+ ion coordination cage, explaining why the D398A mutant GroEL retains only 2% of the wild-type ATPase activity, even though its affinity for ATP is unaffected (40). Furthermore, if Asp398 is prevented from assuming this new active-site position through restriction of domain movements by covalent cross-linking, GroEL is unable to hydrolyze bound ATP (75). These observations reaffirm the conclusion drawn from the fully liganded ATPγ S-bound structure; namely, that binding of ATP to GroEL does not require shifts of the intermediate or apical domains but that subsequent hydrolysis of ATP does.…”

Section: The Asymmetrical Groel-groes-adp Complex Structurementioning

confidence: 99%

STRUCTURE AND FUNCTION IN GroEL-MEDIATED PROTEIN FOLDING

Sigler

Rye

et al. 1998

Annu. Rev. Biochem.

535

387

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Recent structural and biochemical investigations have come together to allow a better understanding of the mechanism of chaperonin (GroEL, Hsp60)-mediated protein folding, the final step in the accurate expression of genetic information. Major, asymmetric conformational changes in the GroEL double toroid accompany binding of ATP and the cochaperonin GroES. When a nonnative polypeptide, bound to one of the GroEL rings, is encapsulated by GroES to form a cis ternary complex, these changes drive the polypeptide into the sequestered cavity and initiate its folding. ATP hydrolysis in the cis ring primes release of the products, and ATP binding in the trans ring then disrupts the cis complex. This process allows the polypeptide to achieve its final native state, if folding was completed, or to recycle to another chaperonin molecule, if the folding process did not result in a form committed to the native state. CONTENTS

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“…Binding of ATP to a GroEL ring drives a large-scale rearrangement of the GroEL subunits, resulting in a dramatic elevation and rotation of the GroEL apical domains away from the central cavity (14,15). GroES binding and substrate folding within the cis complex depend upon these ATP-induced structural rearrangements (5,16,17). However, despite a wealth of structural and biophysical information, precisely how assembly of the GroEL-GroES cis complex leads to substrate protein encapsulation, release, and folding remains poorly understood.…”

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confidence: 99%

Triggering Protein Folding within the GroEL-GroES Complex

Madan¹,

Lin

Rye

2008

Journal of Biological Chemistry

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The folding of many proteins depends on the assistance of chaperonins like GroEL and GroES and involves the enclosure of substrate proteins inside an internal cavity that is formed when GroES binds to GroEL in the presence of ATP. Precisely how assembly of the GroEL-GroES complex leads to substrate protein encapsulation and folding remains poorly understood. Here we use a chemically modified mutant of GroEL (EL43Py) to uncouple substrate protein encapsulation from release and folding. Although EL43Py correctly initiates a substrate protein encapsulation reaction, this mutant stalls in an intermediate allosteric state of the GroEL ring, which is essential for both GroES binding and the forced unfolding of the substrate protein. This intermediate conformation of the GroEL ring possesses simultaneously high affinity for both GroES and non-native substrate protein, thus preventing escape of the substrate protein while GroES binding and substrate protein compaction takes place. Strikingly, assembly of the folding-active GroELGroES complex appears to involve a strategic delay in ATP hydrolysis that is coupled to disassembly of the old, ADP-bound GroEL-GroES complex on the opposite ring.To fold, many essential proteins require the assistance of specialized molecular chaperones known as chaperonins (1). The GroELS chaperonin system of Escherichia coli is one of the best-studied examples of the chaperonin class of molecular chaperones (for review see Refs. 2,3). GroEL is a tetradecamer of fourteen identical 57-kDa subunits arranged into two heptameric rings (4). Each ring contains a large, central, open cavity, and the two rings are stacked back-to-back to create a double toroid. Maximally efficient folding of most proteins that possess a strict dependence on GroEL (so-called stringent substrate proteins) requires that they be encapsulated within a closed chamber formed by a GroEL ring and the separate cochaperonin GroES (5-8). Binding of the GroES lid over a captured substrate protein seals the GroEL cavity and releases the non-native protein into the confined space of the GroEL-GroES chamber (the cis complex). Protein folding proceeds in this isolated space for several seconds until the GroEL-GroES complex is dismantled and the substrate protein, folded or not, is ejected into free solution (5-10).For stringent substrate proteins, ATP is required for the assembly of the GroEL-GroES cis complex and the associated steps of substrate protein encapsulation, release and folding (5, 11-13). Binding of ATP to a GroEL ring drives a large-scale rearrangement of the GroEL subunits, resulting in a dramatic elevation and rotation of the GroEL apical domains away from the central cavity (14,15). GroES binding and substrate folding within the cis complex depend upon these ATP-induced structural rearrangements (5,16,17). However, despite a wealth of structural and biophysical information, precisely how assembly of the GroEL-GroES cis complex leads to substrate protein encapsulation, release, and folding remains poorly understood. Bec...

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GroEL Locked in a Closed Conformation by an Interdomain Cross-link Can Bind ATP and Polypeptide but Cannot Process Further Reaction Steps

Cited by 49 publications

References 43 publications

The Myosin Relay Helix to Converter Interface Remains Intact throughout the Actomyosin ATPase Cycle

The Myosin Relay Helix to Converter Interface Remains Intact throughout the Actomyosin ATPase Cycle

STRUCTURE AND FUNCTION IN GroEL-MEDIATED PROTEIN FOLDING

Triggering Protein Folding within the GroEL-GroES Complex

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