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PubMed2005

Administration of vaccinia virus complement control protein shows significant cognitive improvement in a mild injury model.

Previous studies have shown that traumatic mild brain injury in a rat model is accompanied by breakdown of the blood brain barrier and the accumulation of inflammatory cells. A therapeutic agent, vaccinia virus complement control protein (VCP), inhibits both the classic and the alternative pathways of the complement system and, in so doing, prevents cell death and inflammation. With the use of a rat mild injury model, the effects of VCP on spatial learning and memory were tested. Training in a Morris water maze consisted of a total of 16 trials over a 2-day period before rats were anesthetized and subjected to mild (1.0-1.1 atm) lateral fluid percussion injury (FPI) 3.0 mm lateral to the sagittal suture and 4.5 mm posterior to bregma. Ten microl of VCP (1.7 mg/ml) was injected into the injury site immediately after FPI. Two weeks post-FPI the rats were assessed in the Morris water maze for spatial learning and memory. Neurologic motor function tests were carried out after FPI for 14 consecutive days and again after 28 days. The Morris water maze data show that FPI plus saline-injected rats spent a significantly (P <0.05) larger amount of time in one of the incorrect quadrants than did the FPI plus VCP-injected group. Neurologic evaluations 24 hours postinjury revealed differences in sensorimotor function between groups. The results suggest that in a mild injury model, VCP influences neurologic outcome and offers some enhancement in spatial memory and learning.

Animals↗

PubMed2001

Effect of antioxidants on the apoptosis of CHO cells and production of tissue plasminogen activator in suspension culture.

The effects of antioxidants on the apoptosis of Chinese hamster ovary (CHO) 1-15,500 cells and production of tissue plasminogen activator (tPA) in suspension culture were investigated. After cell growth to 2 x 10(5) cells/ml in Ham's F12 medium containing 10% newborn bovine serum (NBS) in a spinner bottle, CHO cells were maintained for 6 d in Ham's F12 medium containing 0 or 0.4% NBS and 10 mM antioxidants, namely, l-ascorbic acid 2-phosphate (VCP) or the reduced form of glutathione (GSH). The viable cell concentrations at day 6 in the serum-free culture with GSH and in the low-serum culture wiht VCP or GSH were 0.57, 1.04 and 1.69 x 10(5) cells/ml, respectively, while those in the serum-free and low-serum cultures without the antioxidants were only 2.33 and 1.17 x 10(3) cells/ml, respectively. The percentages of apoptotic cells in the serum-free and low-serum cultures with VCP (73.2, 44.6%) and GSH (76.9, 38.6%) measured using a flowcytometer after annexin V-FITC/propidium iodide staining were markedly lower than those in the cultures without antioxidants (96.3, 92.5%). The percentage of cells having a high mitochondrial membrane potential among the viable cells in the cultures with antioxidants determined using a flowcytometer after 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide staining was clearly higher than those in the cultures without the antioxidants. The production of tPA in the serum-free and low-serum media with VCP (0.282, 2.92 mg/l) or GSH (1.01, 1.61 mg/l) was markedly higher than that in the cultures without the antioxidants (0.275, 0.689 mg/l). Consequently, the suppression of apoptosis through the maintenance of the membrane potential of mitochondria by VCP or GSH resulted in a marked increase in tPA production by CHO cells in the serum-free and low-serum cultures.

Journal Article↗

PubMed1991

[Computer-aided visuospatial cognitive-performance test].

The validity of a newly developed computer-aided "Visuospatial Cognitive-Performance Test" (VCP-Test) was examined in two populations, 68 normal elderly volunteers (mean age, 69.5% years) and 25 patients with dementia (mean age, 68.2 years). None of the cases in the two groups had either motor impairment or ataxia. The VCP-Test was a kind of vigilance task in which a subject is required to tap the key corresponding to the local change-over of the image on the computer display as quickly as possible. After 40 trials, consecutive data of performance efficiency were automatically displayed on the LCD display and printed out. A significant negative correlation between the VCP-Test and age in normal elderly population was recognized. Moreover, the relationship between the VCP-Test and the Hasegawa Dementia Scale (HDS), and the Mini-Mental State (MMS) was also significant in patients with dementia. In conclusion, the VCP-Test was considered to be useful for geriatric neurobehavioral functional assessment and for the detection of dementia.

Aged↗

PubMed2000

Effects of adaptation of vestibulo-ocular reflex function on manual target localization.

The goal of the present study was to determine if adaptive modulation of vestibulo-ocular reflex (VOR) function is associated with commensurate alterations in manual target localization. To measure the effects of adapted VOR on manual responses we developed the Vestibular-Contingent Pointing Test (VCP). In the VCP test, subjects pointed to a remembered target following passive whole body rotation in the dark. In the first experiment, subjects performed VCP before and after wearing 0.5X minifying lenses that adaptively attenuate horizontal VOR gain. Results showed that adaptive reduction in horizontal VOR gain was accompanied by a commensurate change in VCP performance. In the second experiment, bilaterally labyrinthine deficient (LD) subjects were tested to confirm that vestibular cues were central to the spatial coding of both eye and hand movements during VCP. LD subjects performed significantly worse than normal subjects. These results demonstrate that adaptive change in VOR can lead to alterations in manual target localization.

Adaptation, Physiological↗

PubMed2003

[Etiology and diagnostic methods in vocal cord paralysis].

INTRODUCTION: The etiology of vocal cord paralysis (VCP) is varied. There is lack of consensus regarding the choice of investigations to be used in the evaluation of VCP. The aim of this study was to establish the etiology, assess the diagnostic methods used in the evaluation, and outline an algorithm for future evaluation of unilateral vocal cord paralysis (UVCP). MATERIAL AND METHODS: Charts of all patients (n = 94) with the diagnostic code of VCP were reviewed, and reexaminations were performed of patients in whom no etiology was found after the initial symptoms. RESULTS: The etiology of UVCP was neoplasm in 34%, surgical trauma in 12%, and miscellaneous causes in 54%. The etiology of bilateral vocal cord paralysis (BVCP) was neoplasm in 24%, surgical trauma in 24%, and miscellaneous causes in 52%. The reexaminations did not reveal any cancer diseases in the patients concerned. The most effective diagnostic method was CT-scanning while the least effective was thyroid scanning. DISCUSSION: Because cancer is a common cause of VCP a thorough evaluation is necessary. For UVCP we recommend history and physical examination, X-ray of the chest, ultrasonography of the neck, and CT-scanning of the superior mediastinum. If these prove negative, panendoscopy should be performed. Workup of patients with idiopathic VCP should include examination, X-ray of the chest at 6-month intervals, and annual CT-scanning for two years.

Aged↗

PubMed2003

[The pathogenesis and treatment exploration of vocal cord paralysis].

OBJECTIVE: To investigate the causes and effective treatment of vocal cord paralysis(VCP). METHOD: Sixty-five patients with VCP in our hospital in recent 10 years were analyzed. All the patients underwent examination by indirect laryngoscope, stroboscopy or electrolaryngostroboscopy, of which, 31 cases were operated by stiring arytenoid cartilage. RESULT: Fourteen cases with VCP were caused by neck and thoracic tumor; Nine cases were caused by infectations; sixteen cases were caused by neck, thoracic, abdominal openations; twelve cases were caused by trachea intubation; Three cases were caused by gastric intubation; Eleven cases were caused without obvious reason. All the symptoms of VCP were cured after treatment in 37 cases. The improvement rate of hoarseness was 58.46%. CONCLUSION: Patients with hoarseness should be routinely examinated by indirect laryngoscope, stroboscopy or electrolaryngostroboscopy. Arytenoid cartilage of patients with VCP should be stired by indirect laryngoscope or direct laryngoscope or self-retaining laryngoscope as early as possible.

Adolescent↗

PubMed1979

The nucleus of the basal optic root in the pigeon: an electron microscope study.

The ultrastructure of the nucleus of the basal optic root in an avian species (Columba livia) was investigated. The ectomamillary nucleus (EMN) in which terminates the basal optic tract reveals three types of neurons: 1) small round neurons bearing a scanty cytoplasm in organelles, 2) medium-sized neurons, spindle-shaped with a dense population of organelles and 3) large multipolar neurons with well developed perikaryal elements. Some of these neurons have their inner plasma-membrane which fuse to make junctional zones alternating between attachment plates and gap junctions. The analysis of the neuropil displays four types of vesicle-containing profiles (VCP), Type I VCP, identified as optic terminals, are numerous (49%), contain round vesicles (500-550 A) and establish Gray type I contacts principally with dendrites. They also participate in serial and triadic arrangements. Type II VCP have lighter hyaloplasm and are less numerous (6,7%). Rounded vesicles (450-500 A) with a clear content synapse also with Gray type I active zones on dendrites. Some of these profiles have the peculiarity of both a chemical and electrical transmission known as mixed synapses. Type III VCP are larger and contain a mixed population of rounded and flattened vesicles which synapse according to Gray type II. Type IV VCP are characterized by a light hyaloplasm where the microtubules are a predominant organelle. Their active zones are also of Gray type II.

Animals↗

PubMed1994

Biochemical characterization of valosin-containing protein, a protein tyrosine kinase substrate in hematopoietic cells.

Engagement of the T cell antigen receptor (TCR) leads to activation of multiple tyrosine kinases and rapid tyrosine phosphorylation of intracellular protein substrates. A number of these substrates have been identified and they include TCR subunits, phospholipase C-gamma 1, p95vav, and ezrin. In a recent study we have demonstrated that VCP (valosin-containing protein) becomes tyrosine phosphorylated upon TCR cross-linking. Analysis of the predicted amino acid sequence of this protein indicates that it is a member of a family of oligomeric proteins containing duplicated domains with predicted ATPase activity. In the current study we determine the site of tyrosine phosphorylation in VCP, demonstrate that murine VCP indeed is an oligomeric ATPase, and show that the tyrosine phosphorylation of the protein has no effect on VCP ATPase activity. Recent evidence suggests that VCP associates with clathrin. A possible role of tyrosine phosphorylation in regulating this protein-protein interaction is discussed.

Adenosine Triphosphatases↗

PubMed2005

Mutant valosin-containing protein causes a novel type of frontotemporal dementia.

Mutations in the valosin-containing protein (VCP) gene on chromosome 9p13-p12 recently have been shown to cause autosomal dominant inclusion body myopathy associated with Paget's disease of the bone and frontotemporal dementia. Here, we report the central nervous system autopsy findings in a 55-year-old German patient with inclusion body myopathy and frontotemporal dementia who harbors a heterozygous R155C missense mutation residing in the N-terminal CDC48 domain of VCP, which is involved in ubiquitin binding. We demonstrate that mutant VCP causes a novel type of frontotemporal dementia characterized by neuronal nuclear inclusions containing ubiquitin and VCP.

Adenosine Triphosphatases↗

PubMed1998

Molecular mimicry of the inflammation modulatory proteins (IMPs) of poxviruses: evasion of the inflammatory response to preserve viral habitat.

Microorganisms encode numerous immunomodulators that resemble, in structure and function, molecules captured over the millennia from their hosts [G. J. Kotwal J. Leukoc. Biol. 62, 415-429]. The vaccinia virus complement control protein (VCP) was the first soluble microbial protein to have a postulated role in the immunomodulation and evasion of host defense [G. J. Kotwal and B. Moss Nature 355, 176-179]. Purified bioactive VCP has been shown to bind to C3 and C4, block the complement cascade at multiple sites [G. J. Kotwal et al. Science 250, 827-830; R. Mckenzie, G. J. Kotwal et al. J. Infect. Dis. 166, 1245-1250] and exhibit a greater potency than the human complement 4b binding protein, C4b-BP [G. J. Kotwal, Am. Biotech. Lab. 9, 76]. The importance of this protein to poxviruses was further demonstrated in rabbits and guinea pigs through the use of recombinant virus lacking an intact DNA coding for VCP [Isaacs, G. J. Kotwal, and B. Moss Proc. Natl. Acad. Sci. 89, 628-672]. Studies in mice have shown that the homolog of VCP in cowpox virus (CPV), referred to as the inflammation modulatory protein (IMP) can, in a mouse model, significantly diminish the specific footpad swelling response [C. G. Miller, S. N. Shchelkunov, and G. J. Kotwal Virol. 229, 126-133]. To determine the precise cellular changes at the site of infection, BALB/c mice were subcutaneously injected (in the backs) with CPV or a recombinant virus lacking IMP, CPV-IMP. Differences in histology were observed by staining the adjoining skin tissue sections with hematoxylin & eosin or by removal of the connective tissue and staining with May-Grunwald-Geimsa. All mice that were injected with the CPV-IMP experienced severe tissue destruction and formation of nodular lesions compared with the mice injected with CPV. Microscopic examination indicated significantly greater cellular infiltration and destruction of skeletal muscle cells in the sections of connective tissue and adjoining skin tissue, respectively, of the mice injected with the CPV-IMP [G. J. Kotwal et al. Mol. Cell. Biochem. in press]. Thus IMP preserves the tissue at the site of infection (viral habitat). In this review, we present evidence for molecular mimicry and evolutionary relationship to other homologs of IMP and discuss their relationships with other IMPs such as the poxviral chemokine and cytokine receptor-like proteins.

Amino Acid Sequence↗

PubMed1997

NMR studies of a viral protein that mimics the regulators of complement activation.

Vaccinia virus complement control protein (VCP) is a 243-residue protein that is similar in sequence to the regulators of complement activation; its role is to defend the virus against attack by the host complement system. A fragment of this protein spanning the two complement protein (CP)-modules (residues 126 to 243) which make up the C-terminal half of VCP has been expressed in Pichia pastoris. A 15N-labelled sample was purified for the purposes of structure determination and measurements of dynamics in solution using NMR. Structures were calculated on the basis of 1767 NMR-derived distance and angle restraints, with a longer than normal high-temperature simulated annealing (SA) protocol which improved convergence. The viral CP-modules are structurally very similar to the 15th and 16th CP-modules of human factor H (fH; average r.m.s.d., for invariant Trp and Cys, four pair-wise comparisons,=1.2 A) but less similar to the fifth CP-module of fH (average r.m.s.d.=2.2 A). In the VCP fragment, the orientation of one module with respect to the other is clearly defined by the experimental data, and T1 measurements are consistent with only limited flexibility at the module-module interface. The r.m.s.d. over all of the 118 residues (backbone atoms) is 0.73 A. The intermodular orientation is better defined than, and significantly different from, that observed in a CP-module pair from fH (re-calculated using the extended SA protocol). In VCP the long axis of the second module is tilted by 59(+/-4) degrees with respect to the first module (50(+/-13) degrees in the fH pair), and twisted with respect to the first module by 22(+/-6) degrees (223(+/-17) degrees in fH). The differences between the human and viral proteins may be rationalised in terms of the lack of hydrogen-bond stabilised secondary structure in the N-terminal portion of fH module 16, and the number and type of amino acid side-chains which make up the interface. A similar intermodular interface may be predicted between the third and fourth module of human C4 binding protein and, probably, between the third and fourth modules of the guinea pig acrosomal matrix protein 67; but the formulation of general rules for predicting the structure of interfaces between CP-modules awaits further experimental data.

Amino Acid Sequence↗

PubMed2003

Vaccinia complement control protein: multi-functional protein and a potential wonder drug.

Vaccinia virus complement control protein (VCP) was one of the first viral molecules demonstrated to have a role in blocking complement and hence in the evasion of host defense. Structurally it is very similar to the human C4b-BP and the other members of complement control protein. Functionally it is most similar to the CR1 protein. VCP blocks both major pathways of complement activation. The crystal structure of VCP was determined a little over a year ago and it is the only known structure of an intact and complete complement control protein. In addition to binding complement, VCP also binds to heparin. These two binding abilities can take place simultaneously and contribute to its many function and to its potential use in several inflammatory diseases, e.g. Alzheimer's disease (AD), CNS injury, xenotransplantation, etc. making it a truly fascinating molecule and potential drug.

Complement Activation↗

PubMed2006

Prognostic value of the VE/VCO2 slope calculated from different time intervals in patients with suspected heart failure.

BACKGROUND: Maximal exercises testing, whether involving cycling- or walking-based protocols, are often not well tolerated in patients with chronic heart failure (CHF). The peak oxygen consumption and the slope of the relation between ventilation (V(E)) and carbon dioxide production (V(CO(2))) are independent predictors of outcome and help risk stratification. The prognostic usefulness of submaximal exercise testing is not clear. The aim of the present study was to assess the prognostic value of the V(E)/V(CO(2)) slope when derived from data acquired from submaximal exercise. METHODS: 394 patients referred with breathlessness and suspected heart failure (74% male) (mean+/-S.D.) age 60+/-12 years; BMI 27+/-5 performed a CPET to determine peak V(O(2)) and the V(E)/V(CO(2)) slope. The V(E)/V(CO(2)) slope was calculated using least squares regression from data acquired from the first 25% of exercise (mean V(E)/V(CO(2)) slope+/-SD; 30.6+/-5.7), 50% (29.6+/-6.9), below the ventilatory compensation point (sub-VCP) (29.9+/-6.8), and all data points (full slope) (32.1+/-7.8). For each measure, patients were divided into quartiles and Kaplan-Meier curves were constructed to determine probability of death after 24 months. The prognostic value of the different classifications was assessed using the chi(2) statistic from the Mantel-Cox log-rank test. RESULTS: During a mean follow-up period of 41+/-19 months, 48 patients died. For the V(E)/V(CO(2)) slope, the log-rank statistic was greatest for the full slope (chi(2)=53.7; P=0.0001), followed by the sub-VCP (chi(2)=45.5; P=0.0001), 50% (chi(2)=41.9; P=0.0001), and 25% (chi(2)= 26.0; P=0.01). The pair-wise log-rank statistic between the fourth and third quartiles was also greatest using the full slope (chi(2)=25.4; P=0.001) followed in order by the sub-VCP (chi(2)=20.1; P=0.001), 50% (chi(2)=19.7; P=0.001), and 25% (chi(2)=14.2; P=0.05). Using the stratified slope measurements entered into a Cox regression analysis using a forward LR stepwise elimination procedure; only the full slope remained significant (P=0.0001). CONCLUSIONS: The V(E)/V(CO(2)) slope should be calculated from all data points to optimise prognostic sensitivity. Data acquired from the first 50% of exercise and below the VCP provide adequate prognostic surrogates in patients who may not be able to perform maximal exercise testing (i.e. in patients with a respiratory exchange ratio<1.10).

Aged↗

PubMed1998

Ethanol is a potent inhibitor of canine cerebrospinal fluid production: an acute and reversible effect.

Neuropathological studies have shown increased cerebral spaces in alcoholics, yet, the effect of ethanol on cerebrospinal fluid (CSF) production is not known. We investigated the effects of ethanol on CSF production measured by ventriculocisternal perfusion (VCP) technique, in two groups (n=10 in each) of anesthetized, paralyzed and mechanically ventilated dogs. In group I, which served as control, VCP was performed with normal mock CSF. Ethanol (150 mg/dl of mock CSF, approximately 33 mM) was added to VCP in group II. Beginning 60 min after the start of VCP, CSF production was measured every 15 min for the next 4 h. In group I, mean (+/-S.D.) value for CSF production was 51+/-10 microliter/min initially and decreased significantly but slightly with time, to the lowest value of 44+/-11 microliter/min at the end of the experiment. In group II, values for CSF production were 41+/-8, 41+/-8, 41+/-8, 43+/-6, 43+/-8, 42+/-6, 42+/-8, 38+/-6, 37+/-6, 36+/-5, 36+/-5, microliter/min, respectively, from 15 to 165 min. These values were invariably significantly lower than their respective mean values in the control group. Furthermore, when ethanol was withdrawn at the trough of CSF production (at 165 min), production significantly increased by about 40%. We conclude that ethanol at a concentration of 150 mg/dl (far below lethal levels) is one of the most potent inhibitory drugs for decreasing CSF production. This effect is short-onset and is fully reversible within 15 min of ethanol withdrawal.

Animals↗

PubMed2002

Fluid distribution measurement between body compartments in primates in disclosing fluid retention during prolonged hypokinesia.

Body fluid homeostasis undergoes significant changes during hypokinesia (diminished movement). Understanding of fluid transfer between body fluid compartments and its regulating mechanisms was the aim of this study for disclosing impaired fluid retention during hypokinesia (HK). Studies were done on 12 male Macaca mulatta (rhesus monkeys) aged three to five years (5.15 to 6.56 kg) during 90 days period of pre-HK and 90 days period of HK. All primates were divided equally into two groups: vivarium control primates (VCP) and hypokinetic primates (HKP). Hypokinetic primates were kept for 90 days in small individual cages that restricted their movements in all directions without hindering food and fluid intakes. Control primates were housed in individual cages without their movements being restricted. Total body fluid (TBF), intracellular fluid volume (IFV) and circulating plasma volume (CPV) decreased significantly (p < 0.05), while extracellular fluid volume (EFV) and interstitial fluid volume (IsFV) decreased significantly (p < 0.05) at the initial seven days of the HK period and after the 7th day increase progressively in HKP compared with their pre-HK values and their respective control values in VCP. Fluid excretion, urinary and plasma sodium (Na) and potassium (K) levels increased significantly (p < 0.05), while fluid intake and fluid retention decreased significantly (p < 0.05) in HKP compared with their pre-HK values and their respective controls in VCP. The measured parameters did not change in VCP compared with their pre-HK values. It was concluded that decreased IFV and CPV may demonstrate fluid depletion, while marked increase of fluid loss despite of fluid depletion may demonstrate impair fluid retention during HK. Dissociation between fluid loss and fluid depletion may demonstrate the presence of reduced fluid retention as the mechanism of development of fluid depletion during HK.

Animals↗

PubMed2001

VCP/p97 in abnormal protein aggregates, cytoplasmic vacuoles, and cell death, phenotypes relevant to neurodegeneration.

Neuronal cell death, abnormal protein aggregates, and cytoplasmic vacuolization are major pathologies observed in many neurodegenerative disorders such as the polyglutamine (polyQ) diseases, prion disease, Alzheimer disease, and the Lewy body diseases, suggesting common mechanisms underlying neurodegeneration. Here, we have identified VCP/p97, a member of the AAA+ family of ATPase proteins, as a polyQ-interacting protein in vitro and in vivo, and report on its characterization. Endogenous VCP co-localized with expanded polyQ (ex-polyQ) aggregates in cultured cells expressing ex-polyQ, with nuclear inclusions in Huntington disease patient brains, and with Lewy bodies in patient samples. Moreover, the expression of VCP mutants with mutations in the 2nd ATP binding domain created cytoplasmic vacuoles, followed by cell death. Very similar vacuoles were also induced by ex-polyQ expression or proteasome inhibitor treatment. These results suggest that VCP functions not only as a recognition factor for abnormally folded proteins but also as a pathological effector for several neurodegenerative phenotypes. VCP may thus be an ideal molecular target for the treatment of neurodegenerative disorders.

Adenosine Triphosphatases↗

PubMed2002

Variola virus immune evasion design: expression of a highly efficient inhibitor of human complement.

Variola virus, the most virulent member of the genus Orthopoxvirus, specifically infects humans and has no other animal reservoir. Variola causes the contagious disease smallpox, which has a 30-40% mortality rate. Conversely, the prototype orthopoxvirus, vaccinia, causes no disease in immunocompetent humans and was used in the global eradication of smallpox, which ended in 1977. However, the threat of smallpox persists because clandestine stockpiles of variola still exist. Although variola and vaccinia share remarkable DNA homology, the strict human tropism of variola suggests that its proteins are better suited than those of vaccinia to overcome the human immune response. Here, we demonstrate the functional advantage of a variola complement regulatory protein over that of its vaccinia homologue. Because authentic variola proteins are not available for study, we molecularly engineered and characterized the smallpox inhibitor of complement enzymes (SPICE), a homologue of a vaccinia virulence factor, vaccinia virus complement control protein (VCP). SPICE is nearly 100-fold more potent than VCP at inactivating human C3b and 6-fold more potent at inactivating C4b. SPICE is also more human complement-specific than is VCP. By inactivating complement components, SPICE serves to inhibit the formation of the C3/C5 convertases necessary for complement-mediated viral clearance. SPICE provides the first evidence that variola proteins are particularly adept at overcoming human immunity, and the decreased function of VCP suggests one reason why the vaccinia virus vaccine was associated with relatively low mortality. Disabling SPICE may be therapeutically useful if smallpox reemerges.

Amino Acid Sequence↗

PubMed1995

Identification of cDNA clones encoding valosin-containing protein and other plant plasma membrane-associated proteins by a general immunoscreening strategy.

An approach was developed for the isolation and characterization of soybean plasma membrane-associated proteins by immunoscreening of a cDNA expression library. An antiserum was raised against purified plasma membrane vesicles. In a differential screening of approximately 500,000 plaque-forming units with the anti-(plasma membrane) serum and DNA probes derived from highly abundant clones isolated in a preliminary screening, 261 clones were selected from approximately 1,200 antiserum-positive plaques. These clones were classified into 40 groups by hybridization analysis and 5'- and 3'-terminal sequencing. By searching nucleic acid and protein sequence data bases, 11 groups of cDNAs were identified, among which valosin-containing protein (VCP), clathrin heavy chain, phospholipase C, and S-adenosylmethionine:delta 24-sterol-C-methyltransferase have not to date been cloned from plants. The remaining 29 groups did not match any current data base entries and may, therefore, represent additional or yet uncharacterized genes. A full-length cDNA encoding the soybean VCP was sequenced. The high level of amino acid identity with vertebrate VCP and yeast CDC48 protein indicates that the soybean protein is a plant homolog of vertebrate VCP and yeast CDC48 protein.

Amino Acid Sequence↗