生物化学技术

Single-molecule enzymology has a longer history than is often supposed, with the first measurements being made as early as 1961. However, the development of new technologies has meant that most of the progress has been made in the last two decades. I review the development of single-molecule enzy ...

Molecular motors perform work in cells by moving in an ATP-dependent manner along filamentous tracks. In vitro, the mechanical action of such motor proteins can be investigated by attaching the molecules to surfaces in the so-called gliding or bead assays. Surface attachment protocols ha ...

Single-molecule measurements of rotation catalyzed by the F1-ATPase or the FoF1 ATP synthase have provided new insights into the molecular mechanisms of the F1 and Fo molecular motors. We recently developed a method to record ATPase-driven rotation of F1 or FoF1 in a manner that solves several t ...

F1-ATPase is the smallest rotary molecular motor ever found. Unidirectional rotation of the γ-shaft is driven by precisely coordinated sequential ATP hydrolysis reactions in three catalytic sites arranged 120� apart in the cylinder. Single-molecule observation allows us to direc ...

Atomic Force Microscopy (AFM) has gained increasing popularity over the years among biophysicists due to its ability to image and to measure pN to nN forces on biologically relevant scales (nm to μm). Continuous technical developments have made AFM capable of nondisruptive, subsecond ima ...

Kinesin motors couple ATP hydrolysis to movement along microtubules, which act both as tracks and as activators of kinesin ATPase activity. Cryo-electron microscopy and image processing enables generation of three-dimensional snapshots of kinesin motors on their tracks at diffe ...

Myosins are mechano-enzymes that convert the chemical energy of ATP hydrolysis into mechanical work. They are involved in diverse biological functions including muscle contraction, cell migration, cell division, hearing, and vision. All myosins have an N-terminal globular domai ...

Recent advances in single-molecule labeling and detection techniques allow high-resolution imaging of the motion of single molecules. Molecular motors are biological machines that convert chemical energy into mechanical work. Myosin Va (MyoVa) is a well-characterized proce ...

Optical tweezers offer the capability to directly observe nanometre displacements and apply piconewton forces to single proteins. This method has been applied to the study of many different biological systems. Optical tweezers have proven to be particularly useful in studying the fi ...

We have developed an enhanced molecular chaperone-based vaccine through rapid isolation of heat-shock protein 70 peptide complexes (Hsp70.PC) after the fusion of tumor and dendritic cells (DCs) (Hsp70.PC-F). In this approach, the tumor antigens are introduced into the antigen-proc ...

Hsp60 (also called Cpn60) is a chaperonin with essential functions for cell physiology and survival. Additionally, its involvement in the pathogenesis of a number of diseases (e.g., some autoimmune disorders and cancer) is becoming evident with new research. For example, the distributi ...

Cell death (in particular, apoptosis and necrosis) is accompanied by appearance of certain hallmarks that are manifested as specific alterations in cellular membranes, cytoplasm, nucleus and mitochondria. Some of those hallmarks are easily detectable in situ and, therefore, they c ...

Heat shock protein 70 (Hsp70) is a powerful chaperone whose expression is induced in response to a wide variety of physiological and environmental insults, including anticancer chemotherapy, thus allowing the cell to survive to lethal conditions. Hsp70 cytoprotective properties m ...

We describe a computational protocol to identify functional modules and pathway relationship of chaperones based on physical interaction data derived from high-throughput proteomic experiments. The protocol first identifies interacting proteins shared by the different ...

Mass spectrometry (MS) is a powerful analytical tool for proteomics research and drug and biomarker discovery. MS enables identification and quantification of known and unknown compounds by revealing their structural and chemical properties. Proper sample preparation for MS-b ...

The use of flow cytometry in heat-shock protein (HSP) research is increasing rapidly due to the high sensitivity and versatility of the technique. The method allows the simultaneous analysis of multiple proteins within numerous cell types in a heterogeneous sample, providing advantag ...

The enzyme-linked immunosorbent assay (ELISA) is an immunological technique which is used to determine the presence or quantity of an antigen within a sample. ELISAs rely on the use of at least one antibody (Ab) specific for the antigen being measured. This antibody is covalently linked to an enzy ...

Stress-inducible heat-shock proteins (HSPs, like HSP70 and HSP27) are molecular chaperones that �protect cells from stress damage by keeping cellular proteins in a folding competent state and preventing them from irreversible aggregation. HSP27 and HSP70 chaperone activities ...

Elevated heat shock protein 27 (Hsp27) expression has been found in a number of tumors, including breast, prostate, gastric, uterine, ovarian, head and neck, and tumor arising from the nervous system and urinary system, and determined to be a predictor of poor clinical outcome. Although the mecha ...

Heat-shock transcription factors (Hsfs) regulate transcription of heat-shock proteins as well as other genes whose promoters contain heat-shock elements. There are at least five Hsfs in mammalian cells, Hsf1, Hsf2, Hsf3, Hsf4, and Hsfy. To understand the physiological roles of Hsf1, Hsf2, ...