Oviductal protease and trypsin treatment enhance sperm–envelope interaction inBufo arenarum coelomic eggs

2006 ◽  
Vol 305A (10) ◽  
pp. 872-882 ◽  
Author(s):  
Ricardo J. Llanos ◽  
Daniel Barrera ◽  
Jorge N. Valz-gianinet ◽  
Dora C. Miceli
Keyword(s):  
Trypsin Treatment

Biosynthesis and Secretion of Serine Peptidase SerP38 from Tenebrio molitor in the Yeast Komagataella kurtzmanii

2020 ◽  
Vol 36 (6) ◽  
pp. 78-86
Author(s):  
P.I. Akentyev ◽  
I.I. Gubaidullin ◽  
N.I. Zhiganov ◽  
V.F. Tereshchenkova ◽  
E.N. Elpidina ◽  
...  
Keyword(s):  
Culture Medium ◽  
Polypeptide Chain ◽  
Tenebrio Molitor ◽  
Enzyme Secretion ◽  
Trypsin Treatment ◽  
Yellow Mealworm ◽  
Serine Peptidase

A strain of the Komagataella kurtzmanii yeast, a producer of recombinant peptidase SerP38 from the yellow mealworm Tenebrio molitor, has been obtained. The level of the pro-enzyme secretion was 20-50 mg/L. It was shown that, during secretion in yeast, the target His6-tagged protein was produced in two forms. One of them was a monomer that was efficiently purified by Ni-NTA chromatography and then activated with trypsin. Another form accumulated in the culture medium as oligomers prone to aggregation in the presence of Ni2+ ions and was not activated by trypsin treatment. Aggregation is likely the result of incorrect folding of the polypeptide chain. Tenebrio molitor, S1 family serine peptidase, SerP38, yeast, Komagataella kurtzmanii, ion-dependent aggregation

Activation of influenza A viruses by trypsin treatment

Virology ◽  
1975 ◽  
Vol 68 (2) ◽  
pp. 426-439 ◽  
Author(s):  
Hans-Dieter Klenk ◽  
Rudolf Rott ◽  
Michaela Orlich ◽  
Jochen Blödorn
Keyword(s):  
Influenza A ◽  
Trypsin Treatment ◽  
Influenza A Viruses

scholarly journals Isolated Epithelial Cells of the Toad Bladder

1968 ◽  
Vol 51 (6) ◽  
pp. 770-784 ◽  
Author(s):  
J. T. Gatzy ◽  
W. O. Berndt
Keyword(s):  
Epithelial Cells ◽  
Connective Tissue ◽  
Divalent Cation ◽  
Toad Bladder ◽  
Trypsin Treatment ◽  
Na Transport ◽  
Isolated Cells ◽  
General Increase ◽  
High Sucrose ◽  
Isolated Cell

Epithelial cells of the toad bladder were disaggregated with EDTA, trypsin, hyaluronidase, or collagenase and were then scraped free of the underlying connective tissue. In most experiments EDTA was complexed with a divalent cation before the tissue was scraped. QOO2, sucrose and inulin spaces, and electrolytes of the isolated cells were measured. Cells disaggregated by collagenase or hyaluronidase consumed O2 at a rate of 4 µl hr-1 dry wt-1. QOO2 was increased 50% by ADH (100 U/liter) or by cyclic 3',5'-AMP (10 mM/liter). Na+-free Ringer's depressed the QOO2 by 40%. The QOO2 of cells prepared by trypsin treatment or by two EDTA methods was depressed by Na+-free Ringer's but was stimulated relatively little by ADH. Two other EDTA protocols produced cells that did not respond to Na+ lack or ADH. The intracellular Na+ and K+ concentrations of collagenase-disaggregated cells were 32 and 117 mEq/kg cell H2O, respectively. Cation concentrations of hyaluronidase cells were similar, but cells that did not respond to ADH had higher intracellular Na+ concentrations. Cells unresponsive to ADH and Na+ lack had high sucrose spaces and low transcellular membrane gradients of Na+, K+, and Cl-. The results suggest that trypsin and EDTA disaggregation damage the active Na+ transport system of the isolated cell. Certain EDTA techniques may also produce a general increase in permeability. Collagenase and hyaluronidase cells appear to function normally.

scholarly journals DISTRIBUTION OF FIBROBLAST SURFACE ANTIGEN

1974 ◽  
Vol 140 (6) ◽  
pp. 1522-1533 ◽  
Author(s):  
Jorma Wartiovaara ◽  
Ewert Linder ◽  
Erkki Ruoslahti ◽  
Antti Vaheri
Keyword(s):  
Surface Antigen ◽  
Rous Sarcoma Virus ◽  
Growth Control ◽  
Trypsin Treatment ◽  
Membrane Processes ◽  
Rous Sarcoma ◽  
Cell Surface Structures ◽  
Sarcoma Virus ◽  
A Cell ◽  
Cell Type Specific

The localization of a cell type-specific, soluble fibroblast surface antigen (SFA) was studied by immunofluorescence and by scanning electron microscopy of the same cells. The antigen had an uneven distribution forming streaks on chick embryo fibroblasts. It was localized to membrane processes and ridges, with a diameter of 50–200 nm. The processes extended from the periphery of the cells to the substratum or to other cells. Trypsin treatment completely removed detectable amounts of SFA. The antigen was detectable within 1 h after trypsin-treated cells were reseeded. The reappearance of SFA correlated with the restoration of membrane processes. Fibroblasts transformed by Rous sarcoma virus (RSV) showed loss of all or most SFA. When normal cells were transformed without subcultivation and trypsinization a fibrillar extracellular network of SFA remained under the transformed fibroblasts while the cells themselves were negative in immunofluorescence. When fibroblasts infected by RSV mutants were transferred to nonpermissive temperature for transformation new SFA was detected within 2 h. These data lead us to propose that loss of stabilizing and anchoring effect of SFA molecules in fibrillar cell surface structures may be critical in altered growth control and malignant transformation.

Membrane-binding properties of filensin, a cytoskeletal protein of the lens fiber cells

1992 ◽  
Vol 103 (3) ◽  
pp. 709-718 ◽  
Author(s):  
M. Brunkener ◽  
S.D. Georgatos
Keyword(s):  
Binding Capacity ◽  
Lens Fiber ◽  
Alkaline Ph ◽  
Trypsin Treatment ◽  
Binding Properties ◽  
Fiber Cells ◽  
Lens Membranes ◽  
Acylated Proteins ◽  
Triton X

Filensin is a 100/110 kDa membrane-associated protein found in lens fiber cells. Previous studies have shown that this protein polymerizes in vitro and binds strongly to vimentin and to another 47 kDa lens membrane protein. Using cosedimentation assays, flotation assays and immunoelectron microscopy, we have examined the properties of purified filensin and measured its binding to lens membranes. Filensin behaves as a ureaextractable, hydrophilic protein which does not partition with Triton X-114 and is not affected by 1 M hydroxylamine at alkaline pH, an agent known to release fatty-acylated proteins from the membrane. Immunoblotting of urea-extracted lens membranes with two different affinity-purified antibodies reveals that, unlike intact filensin, a COOH-terminal filensin degradation product (51 kDa) remains tightly associated with the membranes. Purified filensin binds directly to urea-stripped lens membranes, but not to protein-free vesicles reconstituted from total lens lipids. The binding of filensin is not significantly influenced by the purified 47 kDa protein. Interestingly, the filensin-binding capacity of urea-extracted membranes is increased at least two-fold after trypsin treatment, which removes entirely the 51 kDa peptide from the membranes and presumably unmasks additional filensin-acceptor sites. Consistent with this, filensin binds to trypsinized and non-trypsinized membranes with similar affinities (2 × 10(−7) and 4 × 10(−7) M, respectively). Treatment of the membranes with thrombin, which also eliminates the 51 kDa peptide, does not increase their binding capacity, apparently because filensin-acceptor sites are also destroyed during proteolysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell surface lipids and adhesion. IV. The effects of trypsin on lipid turnover by the plasmalemma

1979 ◽  
Vol 38 (1) ◽  
pp. 283-292
Author(s):  
A.S. Curtis ◽  
O. Hill
Keyword(s):  
Free Energy ◽  
Enzyme Treatment ◽  
Embryonic Chick ◽  
Trypsin Treatment ◽  
Acyltransferase Activity ◽  
Intact Cells ◽  
Coa Ligase ◽  
Surface Lipids ◽  
Lipid Turnover ◽  
Phospholipase A2 Activity

Trypsin treatment of intact cells or isolated plasmalemmae from embryonic chick neural retinae leads to an accumulation of lysophospholipids in the plasmalemmae. Trypsin was used at activities commonly used in cell disaggregation techniques. This accumulation appears to result from the decrease in acyltransferase activity in the plasmalemma produced by enzyme treatment. Plasmalemmal CoA ligase activity is not affected by trypsin treatment. Trypsinization has little effect on plasmalemmal phospholipase A2 activity. These results are discussed in relation to (a) the effects of trypsinization on cell adhesion, and (b) the theory that cells cannot adhere to lecithins because of their fluidity or surface-free-energy values. We propose that the effects of trypsinization on adhesion may in large part be due to the effects on other plasmalemmal proteins. Similarly the inability of cells to adhere to lecithin substrates is simply explained as being due to the lysolecithin that contacting cells release from these substrates.

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