41 research outputs found
Dynamic Proteomics of Individual Cancer Cells in Response to a Drug
Why do seemingly identical cells respond differently to a drug? To address this, we studied the dynamics and variability of the protein response of human cancer cells to a chemotherapy drug, camptothecin. We present a dynamic-proteomics approach that measures the levels and locations of nearly 1000 different endogenously tagged proteins in individual living cells at high temporal resolution. All cells show rapid translocation of proteins specific to the drug mechanism, including the drug target (topoisomerase-1), and slower, wide-ranging temporal waves of protein degradation and accumulation. However, the cells differ in the behavior of a subset of proteins. We identify proteins whose dynamics differ widely between cells, in a way that corresponds to the outcomes—cell death or survival. This opens the way to understanding molecular responses to drugs in individual cells
A Characterization of Scale Invariant Responses in Enzymatic Networks
An ubiquitous property of biological sensory systems is adaptation: a step
increase in stimulus triggers an initial change in a biochemical or
physiological response, followed by a more gradual relaxation toward a basal,
pre-stimulus level. Adaptation helps maintain essential variables within
acceptable bounds and allows organisms to readjust themselves to an optimum and
non-saturating sensitivity range when faced with a prolonged change in their
environment. Recently, it was shown theoretically and experimentally that many
adapting systems, both at the organism and single-cell level, enjoy a
remarkable additional feature: scale invariance, meaning that the initial,
transient behavior remains (approximately) the same even when the background
signal level is scaled. In this work, we set out to investigate under what
conditions a broadly used model of biochemical enzymatic networks will exhibit
scale-invariant behavior. An exhaustive computational study led us to discover
a new property of surprising simplicity and generality, uniform linearizations
with fast output (ULFO), whose validity we show is both necessary and
sufficient for scale invariance of enzymatic networks. Based on this study, we
go on to develop a mathematical explanation of how ULFO results in scale
invariance. Our work provides a surprisingly consistent, simple, and general
framework for understanding this phenomenon, and results in concrete
experimental predictions
Single domain antibodies: promising experimental and therapeutic tools in infection and immunity
Antibodies are important tools for experimental research and medical applications. Most antibodies are composed of two heavy and two light chains. Both chains contribute to the antigen-binding site which is usually flat or concave. In addition to these conventional antibodies, llamas, other camelids, and sharks also produce antibodies composed only of heavy chains. The antigen-binding site of these unusual heavy chain antibodies (hcAbs) is formed only by a single domain, designated VHH in camelid hcAbs and VNAR in shark hcAbs. VHH and VNAR are easily produced as recombinant proteins, designated single domain antibodies (sdAbs) or nanobodies. The CDR3 region of these sdAbs possesses the extraordinary capacity to form long fingerlike extensions that can extend into cavities on antigens, e.g., the active site crevice of enzymes. Other advantageous features of nanobodies include their small size, high solubility, thermal stability, refolding capacity, and good tissue penetration in vivo. Here we review the results of several recent proof-of-principle studies that open the exciting perspective of using sdAbs for modulating immune functions and for targeting toxins and microbes