By mild PAGE method, 11, 11, 7 and 9 chlorophyll_protein complexes were isolated from two species of siphonous green algae (Codium fragile (Sur.) Hariot and Bryopsis corticulans Setch.), green alga (Ulothrix flacca (D...By mild PAGE method, 11, 11, 7 and 9 chlorophyll_protein complexes were isolated from two species of siphonous green algae (Codium fragile (Sur.) Hariot and Bryopsis corticulans Setch.), green alga (Ulothrix flacca (Dillw.) Thur.), and spinach (Spinacia oleracea Mill.), respectively. Apparent molecular weights, Chl a/b ratios, distribution of chlorophyll, absorption spectra, low temperature fluorescence spectra of these complexes were determined, and compared with one another. PSⅠ complexes of two siphonous green algae are larger in apparent molecular weight because of the attachment of relative highly aggregated LHCⅠ. Four isolated light_harvesting complexes of PSⅡ are all siphonaxanthin_Chl a/b_protein complexes, and they are not monomers and oligomers like those in higher plants. Especially, the absence of 730 nm fluorescence in PSⅠ complexes indicates a distinct structure and energy transfer pattern.展开更多
Influence of high light stress on the photosynthesis of flag leaves of indica subspecies (cv. “Shanyou 63', sensitive to photoinhibition) and japonica subspecies (cv. “Wuyujing', resistant to photoin...Influence of high light stress on the photosynthesis of flag leaves of indica subspecies (cv. “Shanyou 63', sensitive to photoinhibition) and japonica subspecies (cv. “Wuyujing', resistant to photoinhibition) of rice ( Oryza sativa L.) was comparatively investigated. In both cultivars of rice, the excitation energy distribution between two photosystems was altered and the excitation energy transfer from light harvesting chlorophyll protein complexes to PSⅡ was inhibited by high light stress. These decreases were more pronounced in indica rice cultivar as compared to japonica one. The analysis of mild SDS_PAGE showed that in indica rice, high light stress almost disaggregated the trimer of light harvesting chlorophyll protein complexes of PSⅡ (LHC Ⅱ 1). The stress reduced the contents of internal antennae chlorophyll protein complexes of PSⅡ (CPa), light harvesting chlorophyll protein of PSⅠ (CPⅠa) and Chl a protein complex of PSⅠ reaction center (CPⅠ) as well as dimer of LHCⅡ (LHCⅡ 2) in indica rice. In japonica subspecies, however, high light stress depressed the contents of LHCⅡ 1, CPa and CPⅠa, but slightly impacted on CPⅠ content. Moreover, the increase in the contents of monomer of LHCⅡ by high light stress was found in both subspecies. In consistent with above results, analysis of polypeptide indicated that the amounts of 27 kD and 25 kD polypeptide of LHCⅡ in particular, as well as that of 21 kD polypeptide of CPⅠa were reduced by high light stress in both subspecies. It was found that, comparing with japonica rice, the stress pronouncedly diminished 43 kD and 47 kD proteins of CPa and 23 kD extrisic protein in indica rice.展开更多
The changes of chlorophyll_protein complexes and photosynthetic activities of chloroplast isolated from lotus ( Nelumbo nucifera Gaertn.) seeds germinating under illumination were studied. SDS PAGE analysis of c...The changes of chlorophyll_protein complexes and photosynthetic activities of chloroplast isolated from lotus ( Nelumbo nucifera Gaertn.) seeds germinating under illumination were studied. SDS PAGE analysis of chlorophyll_protein complexes showed that there was only the light harvesting chlorophyll a/b protein complex from PSⅡ (LHCⅡ) precursor in chloroplast from lotus seeds germinated for 2 to 6 days, while LHC Ⅱ 1, and the chlorophyll_protein complex of PSⅠ (CPⅠ) appeared on the 8th day of germination and PSⅡ reaction center complex appeared later. Studies on the polypeptides composition of the chloroplast revealed the following results: 1) Small amount of the 27 kD polypeptide was synthesized in invisible light; 2) The 30 kD polypeptide existed previously in the plumules of the dry seeds; 3) The amount of the 30 kD polypeptide was more than any other polypeptides before germination and decreased gradually throughout germination, while the 27 kD polypeptide changed in the opposite way; 4) In the process of germination, measurement of the electron transport rate and the fluorescence induction kinetics at room temperature showed that PSⅡ activities and efficiency of primary light energy transformation were only experimentally measurable after 7 days of germination and gradually increased afterwards. At the same time, the chl a/b ratio rose from the lower value to normal; 5) The changes of chloroplast membrane components and its functions are concomitant in concert with that of the ultrastructure of chloroplast membranes during germination, as shown in our earlier work . The results have proved again that a different developmental pathway of chloroplast is likely to exist in the lotus plumules, which might provide an important clue for N. nucifera in having an unique position in the phylogeny of the angiosperm.展开更多
In vertebrates, non-lens βγ-crystallins are widely expressed in various tissues and their functions are not well known. The molecular mechanisms of trefoil factors （TFFs）, which involved in mucosal healing and tum...In vertebrates, non-lens βγ-crystallins are widely expressed in various tissues and their functions are not well known. The molecular mechanisms of trefoil factors （TFFs）, which involved in mucosal healing and tumorigenesis, have remained elusive. βγ-CAT is a novel multifunctional protein complex of non-lens βγ-crystallin and trefoil factor from frog skin secretions. Here we report that βγ-CAT could induce sustained contraction of isolated rabbit aortic rings in dosage （2-35nmol/L） and endothelium dependent manners （P〈0.01 ）. In addition, in situ immunofluorescence indicated that positive TNF-α signals were mainly detected at the endothelial cell layer of βγ-CAT （25nmol/L） treated rings. Furthermore, βγ-CAT induced primary cultured rabbit thoracic aortic endothelial cells （RAECs） rapidly to release TNF-α. After βγ-CAT （25nmol/L） treated for 10 and 30min, the levels of the endothelial cells released TNF-ct were 34.17±5.10 pg/mL and 98.01±4.67 pg/mL （P〈0.01）, respectively. In conclusion, βγ-CAT could induce sustained contraction of isolated aortic rings, and the contractile effect might be partially explained by the release of TNF-α. These findings will give new insight into understanding the functions and physiological roles of non-lens βγ-crystallins and trefoil factors.展开更多
The protecting effect of histidine on the photodamage of pigments and proteins of the isolated PSⅠ particles from the chloroplast of Spinacia oleracea L. during the strong illumination (2 300 μmol·m -2 ...The protecting effect of histidine on the photodamage of pigments and proteins of the isolated PSⅠ particles from the chloroplast of Spinacia oleracea L. during the strong illumination (2 300 μmol·m -2 ·s -1 ) was studied by spectroscopy and SDS_PAGE. The absorbance of PSⅠ particles decreased during the strong illumination treatment, but the decrease would be slowed down in the presence of externally added histidine after 30 min illumination. The decrease of CD (circular dichroism) signal intensities of PSⅠ particles also was slowed down by the added histidine after about 10 min illumination. The retarded protecting effect of the added histidine on the photobleaching of pigments of PSⅠ complexes implied that the mechanisms of photoinhibition of isolated PSⅠ complexes are different from early stage to later stage during the strong illumination treatment. In addition, the added histidine suppressed the decrease of 77 K fluorescence yield of PSⅠ particles during the illumination. SDS_PAGE showed that the added histidine not only protected the reaction center proteins of PSⅠ particles, but also protected other subunits of PSⅠ particles from degradation.展开更多
S-RNase-mediated gametophytic self-incompatibility （GSI） is controlled by a multiallelic S-locus at which two separate genes, the female （pistil） and male （pollen） specificity determinants, are tightly linked. T...S-RNase-mediated gametophytic self-incompatibility （GSI） is controlled by a multiallelic S-locus at which two separate genes, the female （pistil） and male （pollen） specificity determinants, are tightly linked. This review described both the identification of pollen specific F-box genes, SLF/SFBs, in Antirrhinum, Petunia and Prunus species and the demonstration of SLF/SFB as pollen determinant together with their functions in GSI response. Recent studies of how the pollen determinant functions in pollination reaction revealed that pollen determinant interacted with S-RNases in a non-allele-specific manner. It targeted all of the non-self S-RNases for ubiquitination through a functional SCF complex and subsequent degradation via 26S proteasome pathway in compatible reaction. It allows pollen tube to reach into the embryo sac and to finish double fertilization. In incompatible response, the intact self S-RNases were left to function as a cytotoxin that degrades self-pollen tube RNA, resulting in the cessation of pollen tube growth.展开更多
The structural and functional alterations within the PSⅡ membrane from phosphatidylcholine reconstitution and Triton X_100 (TX_100) treatment were studied by using Fourier transform_infrared (FT_IR) spectroscopic tec...The structural and functional alterations within the PSⅡ membrane from phosphatidylcholine reconstitution and Triton X_100 (TX_100) treatment were studied by using Fourier transform_infrared (FT_IR) spectroscopic technique and oxygen electrode. Phosphatidylcholine reconstitution showed no significant effect on the protein secondary structures of PSⅡ membrane but an increase of the rate of PSⅡ_mediated oxygen_evolution. The phosphatidylcholine lipids with different length of acyl chains displayed different capabilities to stimulate oxygen_evolution. In contrast, perturbation of the bilayer lipids by TX_100 resulted in obvious changes of the protein secondary structures within the PSⅡ membrane and in the loss of the PSⅡ_mediated oxygen_evolving activity. The results indicate the importance of membrane integrity in maintaining the stability of the photosynthetic membrane proteins.展开更多
With the identification of more than a dozen novel Hermansky-Pudlak Syndrome （HPS） proteins in vesicle trafficking in higher eukaryotes, a new class of trafficking pathways has been described. It mainly consists of ...With the identification of more than a dozen novel Hermansky-Pudlak Syndrome （HPS） proteins in vesicle trafficking in higher eukaryotes, a new class of trafficking pathways has been described. It mainly consists of three newly-defined protein com- plexes, BLOC-l, -2, and -3. Compelling evidence indicates that these complexes together with two other well-known complexes, AP3 and HOPS, play important roles in endosomal transport. The interactions between these complexes form a network in protein trafficking via endosomes and cytoskeleton. Each node of this network has intra-complex and extra-complex interactions. These complexes are connected by direct interactions between the subunits from different complexes or by indirect interactions through coupling nodes that interact with two or more subunits from different complexes. The dissection of this network facilitates the understanding of a dynamic but elaborate transport machinery in protein/membrane trafficking. The disruption of this network may lead to abnormal trafficking or defective organellar development as described in patients with Hermansky-Pudlak syndrome.展开更多
文摘By mild PAGE method, 11, 11, 7 and 9 chlorophyll_protein complexes were isolated from two species of siphonous green algae (Codium fragile (Sur.) Hariot and Bryopsis corticulans Setch.), green alga (Ulothrix flacca (Dillw.) Thur.), and spinach (Spinacia oleracea Mill.), respectively. Apparent molecular weights, Chl a/b ratios, distribution of chlorophyll, absorption spectra, low temperature fluorescence spectra of these complexes were determined, and compared with one another. PSⅠ complexes of two siphonous green algae are larger in apparent molecular weight because of the attachment of relative highly aggregated LHCⅠ. Four isolated light_harvesting complexes of PSⅡ are all siphonaxanthin_Chl a/b_protein complexes, and they are not monomers and oligomers like those in higher plants. Especially, the absence of 730 nm fluorescence in PSⅠ complexes indicates a distinct structure and energy transfer pattern.
文摘Influence of high light stress on the photosynthesis of flag leaves of indica subspecies (cv. “Shanyou 63', sensitive to photoinhibition) and japonica subspecies (cv. “Wuyujing', resistant to photoinhibition) of rice ( Oryza sativa L.) was comparatively investigated. In both cultivars of rice, the excitation energy distribution between two photosystems was altered and the excitation energy transfer from light harvesting chlorophyll protein complexes to PSⅡ was inhibited by high light stress. These decreases were more pronounced in indica rice cultivar as compared to japonica one. The analysis of mild SDS_PAGE showed that in indica rice, high light stress almost disaggregated the trimer of light harvesting chlorophyll protein complexes of PSⅡ (LHC Ⅱ 1). The stress reduced the contents of internal antennae chlorophyll protein complexes of PSⅡ (CPa), light harvesting chlorophyll protein of PSⅠ (CPⅠa) and Chl a protein complex of PSⅠ reaction center (CPⅠ) as well as dimer of LHCⅡ (LHCⅡ 2) in indica rice. In japonica subspecies, however, high light stress depressed the contents of LHCⅡ 1, CPa and CPⅠa, but slightly impacted on CPⅠ content. Moreover, the increase in the contents of monomer of LHCⅡ by high light stress was found in both subspecies. In consistent with above results, analysis of polypeptide indicated that the amounts of 27 kD and 25 kD polypeptide of LHCⅡ in particular, as well as that of 21 kD polypeptide of CPⅠa were reduced by high light stress in both subspecies. It was found that, comparing with japonica rice, the stress pronouncedly diminished 43 kD and 47 kD proteins of CPa and 23 kD extrisic protein in indica rice.
文摘The changes of chlorophyll_protein complexes and photosynthetic activities of chloroplast isolated from lotus ( Nelumbo nucifera Gaertn.) seeds germinating under illumination were studied. SDS PAGE analysis of chlorophyll_protein complexes showed that there was only the light harvesting chlorophyll a/b protein complex from PSⅡ (LHCⅡ) precursor in chloroplast from lotus seeds germinated for 2 to 6 days, while LHC Ⅱ 1, and the chlorophyll_protein complex of PSⅠ (CPⅠ) appeared on the 8th day of germination and PSⅡ reaction center complex appeared later. Studies on the polypeptides composition of the chloroplast revealed the following results: 1) Small amount of the 27 kD polypeptide was synthesized in invisible light; 2) The 30 kD polypeptide existed previously in the plumules of the dry seeds; 3) The amount of the 30 kD polypeptide was more than any other polypeptides before germination and decreased gradually throughout germination, while the 27 kD polypeptide changed in the opposite way; 4) In the process of germination, measurement of the electron transport rate and the fluorescence induction kinetics at room temperature showed that PSⅡ activities and efficiency of primary light energy transformation were only experimentally measurable after 7 days of germination and gradually increased afterwards. At the same time, the chl a/b ratio rose from the lower value to normal; 5) The changes of chloroplast membrane components and its functions are concomitant in concert with that of the ultrastructure of chloroplast membranes during germination, as shown in our earlier work . The results have proved again that a different developmental pathway of chloroplast is likely to exist in the lotus plumules, which might provide an important clue for N. nucifera in having an unique position in the phylogeny of the angiosperm.
基金National Natural Science Foundation (30630014, 30570359)The grant of "Key Research Direction-KSCX2-YW-R-088" from Chinese Academy of Sciences~~
文摘In vertebrates, non-lens βγ-crystallins are widely expressed in various tissues and their functions are not well known. The molecular mechanisms of trefoil factors （TFFs）, which involved in mucosal healing and tumorigenesis, have remained elusive. βγ-CAT is a novel multifunctional protein complex of non-lens βγ-crystallin and trefoil factor from frog skin secretions. Here we report that βγ-CAT could induce sustained contraction of isolated rabbit aortic rings in dosage （2-35nmol/L） and endothelium dependent manners （P〈0.01 ）. In addition, in situ immunofluorescence indicated that positive TNF-α signals were mainly detected at the endothelial cell layer of βγ-CAT （25nmol/L） treated rings. Furthermore, βγ-CAT induced primary cultured rabbit thoracic aortic endothelial cells （RAECs） rapidly to release TNF-α. After βγ-CAT （25nmol/L） treated for 10 and 30min, the levels of the endothelial cells released TNF-ct were 34.17±5.10 pg/mL and 98.01±4.67 pg/mL （P〈0.01）, respectively. In conclusion, βγ-CAT could induce sustained contraction of isolated aortic rings, and the contractile effect might be partially explained by the release of TNF-α. These findings will give new insight into understanding the functions and physiological roles of non-lens βγ-crystallins and trefoil factors.
文摘The protecting effect of histidine on the photodamage of pigments and proteins of the isolated PSⅠ particles from the chloroplast of Spinacia oleracea L. during the strong illumination (2 300 μmol·m -2 ·s -1 ) was studied by spectroscopy and SDS_PAGE. The absorbance of PSⅠ particles decreased during the strong illumination treatment, but the decrease would be slowed down in the presence of externally added histidine after 30 min illumination. The decrease of CD (circular dichroism) signal intensities of PSⅠ particles also was slowed down by the added histidine after about 10 min illumination. The retarded protecting effect of the added histidine on the photobleaching of pigments of PSⅠ complexes implied that the mechanisms of photoinhibition of isolated PSⅠ complexes are different from early stage to later stage during the strong illumination treatment. In addition, the added histidine suppressed the decrease of 77 K fluorescence yield of PSⅠ particles during the illumination. SDS_PAGE showed that the added histidine not only protected the reaction center proteins of PSⅠ particles, but also protected other subunits of PSⅠ particles from degradation.
基金This work was supported by grants from Three Founda-tions of Hunan Province (00JZY2155) and International Cooperation Project
文摘S-RNase-mediated gametophytic self-incompatibility （GSI） is controlled by a multiallelic S-locus at which two separate genes, the female （pistil） and male （pollen） specificity determinants, are tightly linked. This review described both the identification of pollen specific F-box genes, SLF/SFBs, in Antirrhinum, Petunia and Prunus species and the demonstration of SLF/SFB as pollen determinant together with their functions in GSI response. Recent studies of how the pollen determinant functions in pollination reaction revealed that pollen determinant interacted with S-RNases in a non-allele-specific manner. It targeted all of the non-self S-RNases for ubiquitination through a functional SCF complex and subsequent degradation via 26S proteasome pathway in compatible reaction. It allows pollen tube to reach into the embryo sac and to finish double fertilization. In incompatible response, the intact self S-RNases were left to function as a cytotoxin that degrades self-pollen tube RNA, resulting in the cessation of pollen tube growth.
文摘The structural and functional alterations within the PSⅡ membrane from phosphatidylcholine reconstitution and Triton X_100 (TX_100) treatment were studied by using Fourier transform_infrared (FT_IR) spectroscopic technique and oxygen electrode. Phosphatidylcholine reconstitution showed no significant effect on the protein secondary structures of PSⅡ membrane but an increase of the rate of PSⅡ_mediated oxygen_evolution. The phosphatidylcholine lipids with different length of acyl chains displayed different capabilities to stimulate oxygen_evolution. In contrast, perturbation of the bilayer lipids by TX_100 resulted in obvious changes of the protein secondary structures within the PSⅡ membrane and in the loss of the PSⅡ_mediated oxygen_evolving activity. The results indicate the importance of membrane integrity in maintaining the stability of the photosynthetic membrane proteins.
基金This work was supported in part by the National Science Fund for Distinguished Young Scholars (No. 30525007)National Basic Research Program of China (No. 2006CB504103+1 种基金 No. 2006CB500704)Hi-Tech Research and Development Program of China (No. 2006AA02Z322)
文摘With the identification of more than a dozen novel Hermansky-Pudlak Syndrome （HPS） proteins in vesicle trafficking in higher eukaryotes, a new class of trafficking pathways has been described. It mainly consists of three newly-defined protein com- plexes, BLOC-l, -2, and -3. Compelling evidence indicates that these complexes together with two other well-known complexes, AP3 and HOPS, play important roles in endosomal transport. The interactions between these complexes form a network in protein trafficking via endosomes and cytoskeleton. Each node of this network has intra-complex and extra-complex interactions. These complexes are connected by direct interactions between the subunits from different complexes or by indirect interactions through coupling nodes that interact with two or more subunits from different complexes. The dissection of this network facilitates the understanding of a dynamic but elaborate transport machinery in protein/membrane trafficking. The disruption of this network may lead to abnormal trafficking or defective organellar development as described in patients with Hermansky-Pudlak syndrome.
