Misfolding or unfolding of polypeptides can occur as a consequence of environmental stress and spontaneous mutation. The abundance of general chaperones and proteases suggests that cells distinguish between proteins that can be refolded and "hopeless" cases fated to enter the proteolytic pathway. The mechanisms controlling this key metabolic decision are not well understood. We show here that the widely conserved heat shock protein DegP (HtrA) has both general molecular chaperone and proteolytic activities. The chaperone function dominates at low temperatures, while the proteolytic activity is present at elevated temperatures. These results show that a single cellular factor can switch between two key pathways, controlling protein stability and turnover. Implications of this finding for intracellular protein metabolism are discussed.

Bispecific antibodies are on the cusp of coming of age as therapeutics more than half a century after they were first described. Two bispecific antibodies, catumaxomab (Removab(®), anti-EpCAM×anti-CD3) and blinatumomab (Blincyto(®), anti-CD19×anti-CD3) are approved for therapy, and >30 additional bispecific antibodies are currently in clinical development. Many of these investigational bispecific antibody drugs are designed to retarget T cells to kill tumor cells, whereas most others are intended to interact with two different disease mediators such as cell surface receptors, soluble ligands and other proteins. The modular architecture of antibodies has been exploited to create more than 60 different bispecific antibody formats. These formats vary in many ways including their molecular weight, number of antigen-binding sites, spatial relationship between different binding sites, valency for each antigen, ability to support secondary immune functions and pharmacokinetic half-life. These diverse formats provide great opportunity to tailor the design of bispecific antibodies to match the proposed mechanisms of action and the intended clinical application.

Aggregation of proteins containing polyglutamine (polyQ) expansions characterizes many neurodegenerative disorders, including Huntington’s disease. Molecular chaperones modulate Huntingtin (Htt) aggregation and toxicity by an ill-defined mechanism. Here we determine how the chaperonin TRiC suppresses Htt aggregation. Surprisingly, TRiC does not physically block the polyQ tract itself, but rather sequesters a short Htt sequence element N-terminal to the polyQ tract, that promotes the amyloidogenic conformation. The residues of this amyloid-promoting element essential for rapid Htt aggregation are directly bound by TRiC. Our findings illustrate how molecular chaperones, which recognize hydrophobic determinants, can prevent aggregation of polar polyQ tracts associated with neurodegenerative diseases. The observation that the switch of polyQ tracts to an amyloidogenic conformation is accelerated by short endogenous sequence elements provides a novel target for therapeutic strategies to inhibit aggregation.

Chaperonins are key components of the cellular chaperone machinery. These large, cylindrical complexes contain a central cavity that binds to unfolded polypeptides and sequesters them from the cellular environment. Substrate folding then occurs in this central cavity in an ATP-dependent manner. The eukaryotic chaperonin TCP-1 ring complex (TRiC, also called CCT) is indispensable for cell survival because the folding of an essential subset of cytosolic proteins requires TRiC, and this function cannot be substituted by other chaperones. This specificity indicates that TRiC has evolved structural and mechanistic features that distinguish it from other chaperones. Although knowledge of this unique complex is in its infancy, we review recent advances that open the way to understanding the secrets of its folding chamber.Protein misfolding has been implicated in several human diseases that have both systemic and neurological implications, including 'conformational diseases' such as Huntington's and Parkinson's that are characterized by the accumulation of toxic protein aggregates [1]. Although small proteins with simple chain topologies can fold spontaneously, the vast majority of cellular proteins is unable to reach its native state without the assistance of elaborate cellular machinery composed of proteins known as molecular chaperones (for review, see Refs [1-4]). A complete understanding of chaperone-assisted protein folding in the cell would be an intellectual tour de force that might, eventually, lead to effective treatments for these diseases.In the cell, protein folding faces additional challenges compared with the refolding of proteins in solution [1,2,4]. For example, in vivo, the linear polypeptide chain emerges vectorially into the cytosol during synthesis on ribosomes. Because the information for the native state is encoded by the entire amino acid sequence, the nascent polypeptide chain is unable to fold stably until fully synthesized, but it exposes hydrophobic sequences into the crowded cellular milieu [4]. Details of how newly translated proteins navigate toward a final, fully functional, folded structure in vivo are not entirely understood, but it is clear that exposed hydrophobic surfaces can contribute to misfolding and aggregation. Accordingly, all cellular compartments contain many structurally and functionally distinct classes of chaperones that vary in size and complexity, ranging from those that bind only to misfolded polypeptides and prevent their aggregation to those that recognize specific classes of proteins and facilitate their folding to the native state in an energy-dependent manner [1][2][3]. In cells, these different classes of chaperones work together to form elaborate, cooperative networks that ensure the correct folding of newly translated proteins; they also ensure that potentially damaging misfolded polypeptides are cleared from the cell [5].

Misfolding and aggregation of proteins containing expanded polyglutamine repeats underlie Huntington's disease and other neurodegenerative disorders 1 . Here, we show that the heterooligomeric chaperonin TRiC (also known as CCT) physically interacts with polyglutamine-expanded variants of huntingtin (Htt) and effectively inhibits their aggregation. Depletion of TRiC enhances polyglutamine aggregation in yeast and mammalian cells. Conversely, overexpression of a single TRiC subunit, CCT1, is sufficient to remodel Htt-aggregate morphology in vivo and in vitro, and reduces Htt-induced toxicity in neuronal cells. Because TRiC acts during de novo protein biogenesis 2 , this chaperonin may have an early role preventing Htt access to pathogenic conformations. Based on the specificity of the Htt-CCT1 interaction, the CCT1 substrate-binding domain may provide a versatile scaffold for therapeutic inhibitors of neurodegenerative disease.Late-onset neurodegenerative diseases are often associated with the accumulation of insoluble amyloid aggregates in neurons 3 . In many cases, such as spinocerebellar ataxia and Huntington's disease, aggregation is associated with expanded polyglutamine (polyQ) tracts in the disease gene, usually beyond a critical threshold of approximately 40 glutamine repeats 1 . Because polyQ disease proteins are the main aggregate component in affected neurons 4 , and glutamine tract length correlates with both aggregation propensity and age of onset of disease, it seems that toxic conformations of the polyQ-expanded proteins are directly responsible for neuronal dysfunction and death 1,5 .Recent studies suggest that the age-dependent accumulation of protein aggregates in neurodegenerative diseases reflects the progressive inability of the cellular quality control machinery to recognize and eliminate potentially toxic conformations. Molecular chaperones, which selectively bind non-native proteins and facilitate their folding or degradation 6,7 , have been shown to modulate aggregation and toxicity in neurodegenerative disease models. In particular, overexpression studies have demonstrated that the chaperone Hsp70 and its cofactors, such as Hsp40, can remodel polyQ aggregates and alleviate the toxicity of polyQ aggregation 4 . Although these studies establish a role for chaperones in modulating polyQ aggregation, the chaperones that normally interact with pathogenic polyQ conformations and 3Correspondence should be addressed to J.F. (jfrydman@stanford.edu).