Isolation and Biochemical Characteristics of the Nuclear Envelope

Zbarsky, I. B.; Perevoshchikova, K. A.; Delektorskaya, L. N.; Delektorsky, V V

doi:10.1038/221257a0

Cited by 94 publications

(20 citation statements)

References 8 publications

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“…Attempts to separate inner from outer nuclear membrane by fractionation have also been mentioned in the literature, but the results published do not yet permit one to conclude that the separation was adequate (Zbarsky et al, 1967;Smith et al, 1969;Mizuno et al, 1971a, b;Zbarsky, 1972b). In fact, during most fragmentation treatments the nuclear envelope preferentially breaks into vesicles comprising both inner and outer membrane material, rather than into sheets of either the inner or the outer membrane.…”

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confidence: 99%

“…Methods for preparing nuclear membrane fractions have been reported for various plant tissues such as onion root tip or leaves (Franke, 1966b), for the macronuclei of the ciliate, Tetrahymena pyriformis (Franke, 1967 a, b;Eckert, 1972), for mouse, rat, rabbit, and pig liver (Franke, 1967a, b;Zbarsky et al, 1967Zbarsky et al, , 1969Bornens, 1968;Kashnig and Kasper, 1969;Berezney et al, 1970Franke et al, 1970;Kartenbeck et al, 1971Kartenbeck et al, , 1973Agutter, 1972;Kay et al, 1972;Monneron et al, 1972;Price et al, 1972), for avian erythrocytes (Harris and Brown, 1971;Zentgraf et al, 1971) for rat and calf thymus (Matsuura and U eda, 1972;J arasch et al, 1973), for rat prostate gland (Moore and Wilson, 1972), and for a series of tumor cells (Zbarsky et al, 1967Comes and Franke, 1970;Fakan et al, 1972;Green and Dobrjansky, 1972;Price et al, 1972). The variety of procedures reported has been reviewed in detail by Zbarsky (1972b) and .…”

Section: J\:\mentioning

confidence: 99%

“…(b) Mass isolations of nuclear membrane material usually start from a purified fraction of isolated nuclei. The nuclei are then fragmented and/ or extracted with combinations of diverse treatments: (i) rapid chromatin swelling in hypotonic solutions (Franke 1966a(Franke , b, 1967aZbarsky et al, 1967Zbarsky et al, , 1969Franke and Kartenbeck, 1969;Price et al, 1972; reviewed by Zbarsky, 1972a, b); ( ii) vigorous homogenization, shearing and sonication (Franke, 1966a(Franke, , b, 1967aBornens, 1968;Kashnig and Kasper, 1969;Zbarsky et al, 1969;Comes and Franke, 1970;Harris and Agutter, 1970;Harris and Brown, 1971;Zentgraf et al, 1971;Agutter, 1972;Fakan et al, 1972;Moore and Wilson, 1972;J arasch et al, 1973); (iii) limited digestion of the chromatin with deoxyribonuclease Kay et al, 1971Kay et al, , 1972Zentgraf et al, 1971;Matsuura and Ueda, 1972); (iv) destabilization with chelating agents such as citrate (Bornens, 1968;Kashnig and Kasper, 1969) or in high salt concentrations Franke et al, 1970;ZentFig. 2 Special nucleocytoplasmic separation zone of the primary (giant) nucleus in the rhizoid of the green alga, Acetabularia mediterranea.…”

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Structures and Functions of the Nuclear Envelope

Franke¹,

Scheer²

1974

The Cell Nucleus

153

150

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Structures and Functions of the Nuclear Envelope

Franke¹,

Scheer²

1974

The Cell Nucleus

153

150

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“…Nu-THE JOUENAL OF CELL BIOLOGY . VoLumE 46, 1970 • pages [379][380][381][382][383][384][385][386][387][388][389][390][391][392][393][394][395] clear envelope fragments were isolated from mammalian liver first by the method of Franke (26,27) which uses a combination of hypotonic shock and sonication for the nuclear disruption (15,(26)(27)(28)(29)95) and, recently, by other authors employing similar methods (9, 43,97) . Other attempts for obtaining nuclear membrane material from diverse mammalian cells, such as HeLa cells, calf thymus, and rat liver, involved the use of detergents such as deoxycholate and Triton X-100 (4, 75,92,93) which obviously can cause considerable membrane dissociation and thus progressive loss of membrane material, especially in the outer nuclear membrane (40), and consequently have a very limited range of application .…”

Section: Introductionmentioning

confidence: 99%

Nuclear Membranes From Mammalian Liver

Franke

Deumling

Ermen

et al. 1970

The Journal of Cell Biology

190

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Nuclear membranes were isolated from rat and pig liver by sonication of highly purified nuclear fractions and subsequent removal of adhering nucleoproteins in a high salt medium . The fractions were examined in the electron microscope by both negative staining and thin sectioning techniques and were found to consist of nuclear envelope fragments of widely varying sizes. Nuclear pore complex constituents still could frequently be recognized . The chemical composition of the nuclear membrane fractions was determined and compared with those of microsomal fractions prepared in parallel . For total nuclei as well as for nuclear membranes and microsomes, various enzyme activities were studied . The results indicate that a similarity exists between both fractions of cytomembranes, nuclear envelope, and endoplasmic reticulum, with respect to their RNA : protein ratio and their content of polar and nonpolar lipids . Both membranous fractions had many proteins in common including some membrane-bound enzymes . Activities in Mg-ATPase and the two examined cytochrome reductases were of the same order of magnitude . The content of cytochrome b 5 as well as of P-450 was markedly lower in the nuclear membranes . The nuclear membranes were found to have a higher buoyant density and to be richer in protein . The glucose-6-phosphatase and Na-K-ATPase activities in the nuclear membrane fraction were very low. In the gel electrophoresis, in addition to many common protein bands, some characteristic ones for either microsomal or nuclear membranous material were detected . Significant small amounts of DNA and RNA were found to remain closely associated with the nuclear envelope fragments . Our findings indicate that nuclear and endoplasmic reticulum membranes which are known to be in morphological continuity have, besides a far-reaching similarity, some characteristic differences .

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“…Some workers who have isolated nuclear enve-lopes (Monneron et al ., 1972 ;Franke et al ., 1970 ;Berezney et al ., 1970) did not determine whether or not these envelopes contained sialic acid, but Zbarsky et al . (1969) reported that nuclear envelopes from a variety of sources contained 0 .08-0 .1 % sialic acid .…”

Section: Discussionmentioning

confidence: 99%

Molecules at the External Nuclear Surface

Bosmann

1973

The Journal of Cell Biology

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The molecules occurring as terminal residues on the external surfaces of nuclei prepared from rat liver by either sucrose-CaC12 or citric acid methods and nucleoli derived from the sucrose-CaC1 2 nuclei were studied chemically and electrokinetically . In 0.0145 M NaCl, 4 .5 0 /,) sorbitol, and 0 .6 mM NaHCO3 with pH 7 .2 t 0 .1 at 25°C, the sucrose-CaC12 nuclei had an electrophoretic mobility of -1 .92 ,um/s/V/cm, the citric acid nuclei, -1 .63 µm/s/V/cm, and the nucleoli, -2 .53 µm/s/V/cm . The citric acid nuclei and the nucleoli contained no measurable sialic acid . The sucrose-CaC12 nuclei contained 0.7 pmol of sialic acid/mg nuclear protein ; this was essentially located in the nuclear envelope . Treatment of these nuclei with 50 µg neuraminidase/mg protein resulted in release of 0 .63 pmol of sialic acid/mg nuclear protein ; treatment with 1 % trypsin caused release of 0 .39 nmol of the sialic acid/mg nuclear protein . The pH-mobility curves for the particles indicated the sucrose-CaC12 nuclei surface had an acid-dissociable group of pK -2 .7 while the pK for the nucleoli was considerably lower . Nucleoli treated with 50 µg neuraminidase/mg particle protein had a mobility of -2 .53 Am/s/V/cm while sucrose-CaC12 nuclei similarly treated had a mobility of -1 .41 µm/s/V/cm . Hyaluronidase at 50 µg/mg protein had no effect on nucleoli mobility but decreased the sucrose-CaC12 nuclei mobility to -1 .79 M.m/s/V/cm . Trypsin at 1 % elevated the electrophoretic mobility of the sucrose-CaC12 nuclei slightly but decreased the mobility of the nucleoli to -2 .09 µm/s/V/cm . DNase at 50 Ag/mg protein had no effect on the mobility of the isolated sucrose-CaC12 nuclei but decreased the electrophoretic mobility of the nucleoli to -1 .21 µm/s/V/cm . RNase at 50 µg/mg protein also had no effect on the electrophoretic mobility of the sucrose-CaC12 nuclei but decreased the nucleoli mobility to -2 .10 µm/s/V/cm . Concanavalin A at 50 Ag/mg protein did not alter the nucleoli electrophoretic mobility but decreased the sucrose-CaC1 2 nuclei electrophoretic mobility to -1 .64 µm/s/V/cm . The results are interpreted to mean that the sucrose-CaC1 2 nuclear external surface contains terminal sialic acid residues in trypsin-sensitive glycoproteins, contains small amounts of hyaluronic acid, is completely devoid of nucleic acids, and binds concanavalin A . The nucleolus surface is interpreted to contain a complex made up of protein, RNA, and primarily DNA, to be devoid of sialic acid and hyaluronic acid, and not to bind concanavalin A.

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Isolation and Biochemical Characteristics of the Nuclear Envelope

Cited by 94 publications

References 8 publications

Structures and Functions of the Nuclear Envelope

Structures and Functions of the Nuclear Envelope

Nuclear Membranes From Mammalian Liver

Molecules at the External Nuclear Surface

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