Heat-shock protein 40 (Hsp40) enables Hsp70 to play critical roles in a number of cellular processes, such as protein folding, assembly, degradation and translocation in vivo. Hsp40 recognizes and binds non-native polypeptides and delivers them to Hsp70. Then Hsp40 stimulates the ATPase activity of Hsp70 to fold the polypeptides. By using yeast Hsp40 Sis1 and yeast Hsp70 Ssa1 as our model proteins, we found that the Sis1 peptide-binding fragment interacts directly with the full-length Ssa1 in vitro. Further studies showed that the C-terminal lid domain of Ssa1 could interact with Sis1 peptide-binding domain physically in vitro. The Sis1 peptide-binding fragment forms a stable complex with the Ssa1 C-terminal lid domain in solution. The interactions between these two proteins appear to be charge–charge interactions because high-ionic-strength buffer can dissociate the complex. Further mapping studies showed that the Sis1 peptide-binding fragment binds the extreme C-terminal 15 amino acid residues of Ssa1. A flexible glycine-rich region is followed by these 15 residues in the Ssa1 primary sequence. Atomic force microscopy of the Sis1–Ssa1 complex showed that only one end of the Ssa1 lid domain binds the Sis1 peptide-binding-fragment dimer at the upper level of the huge groove within the Sis1 dimer. Based on the data, we propose an ‘anchoring and docking’ model to illustrate the mechanisms by which Hsp40 interacts with Hsp70 and delivers the non-native polypeptide to Hsp70.

The mouse tumor cell S180 and human liver carcinoma cell SMC 7721 cells were first treated with R-PE and its subunits (alpha, beta, gamma subunits), then irradiated with Argon laser (496 nm, 28.8 J/cm2). Survival rate was measured by MTT method. In order to compare the phototoxicity in normal cells, the mouse marrow cells were treated with photofrin II and beta-subunit, irradiated with 45 J/cm2 of light; survival rate was also measured by MTT method. The result showed that R-PE subunits had better PDT effect on s180 cells than R-PE and lower phototoxicity in marrow cells than photofrin II. Flow cytometric analysis showed that PDT results in a growth inhibition and a G0-G1 cell cycle arrest in SMC 7721 cells. The tumor cells inhibited by PDT in vivo were morphologically observed by TEM, the tumor cell death was due to the occlusion of tumor blood vessels and inducement of cell programmed death in nuclei. Therefore, with the advantage in special fluorescence activity, low molecular weight, good light absorbent character and weak phototoxicity, R-PE subunit is an attractive option for improving the selectivity of PDT.

The concentration of trace gases in the atmospheric environment is extremely low, but it has a great impact on the living environment of organisms. Photoacoustic spectroscopy has attracted extensive attention in the field of trace gas detection because of its high sensitivity, good selectivity, and fast response. As the core of a photoacoustic detection setup, the photoacoustic cell has a significant impact on detection performance. To improve detection sensitivity, a sphere-tube coupled photoacoustic cell (STPAC) was developed, which was mainly composed of a diffuse-reflective sphere and an acoustic resonance tube. Modulated light was reflected multiple times in the sphere to increase optical path, and photoacoustic (PA) signals were further amplified by the tube. Based on STPAC, a PA gas detection setup was built with a laser diode (LD) at 450 nm as the light source. The experimental results showed that the minimum detection limit (noise equivalent concentration, NEC) of NO2 was ~0.7 parts per billion (ppb). Compared with the T-type PA cell (TPAC) in which the modulated light passed through the sphere, the signal-to-noise ratio of STPAC was increased by an order of magnitude at the same concentration of the NO2 sample.