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K Wang

Publications and source records attributed to K Wang.

At least 541 records · Page 30Linked to original sources
PubMed1988

Titin: quantitative mass measurements by scanning transmission electron microscopy and structural implications for the sarcomere matrix of skeletal muscle.

Scanning transmission electron microscopy has been used to investigate mass and linear mass density of native titin-2, a large soluble fragment of intact titin, from rabbit skeletal muscle. Dark field images of unstained, freeze-dried titin-2 appeared as either compact globules or looser and larger balls of string. Direct mass measurements indicated that the compact forms have an average mass of 2.40 +/- 0.50 x 10(6) Da. The mass to length ratio, determined from well-spread portions of titin strands (3-5 nm wide) from the ball of string forms, averaged 2.7 +/- 0.9 kDa/nm. Thus a single native intact titin molecule has a calculated contour length of well above approximately 1 micron, sufficient to span unidirectionally between the Z line and M line region in a resting-length sarcomere.

Animals↗
PubMed1988

Giant polypeptides of skeletal muscle titin: sedimentation equilibrium in guanidine hydrochloride.

Titin has been purified from rabbit skeletal myofibrils in guanidine hydrochloride by gel filtration on Sephacryl S-1000. The sedimentation coefficients of denatured titin in 6 M guanidine hydrochloride are highly concentration dependent and follow the equation: 1/S = 0.241 + 0.302 A280. Equilibrium sedimentation studies indicate that the average molecular weight of titin polypeptides is 2.4 to 2.6 X 10(6).

Animals↗
PubMed1988

Sarcomere matrix of striated muscle: in vivo phosphorylation of titin and nebulin in mouse diaphragm muscle.

Titin and nebulin are two major protein components of a cytoskeletal matrix that coexists with thick and thin filaments within the sarcomere of a wide range of striated muscles. Purified titin and nebulin from mouse diaphragm muscle are similar in size, in relative abundance, and in amino acid composition to analogous proteins from other mammals or avians. Phosphate analysis of these nucleic-acid-free proteins indicated that both proteins contain substantial amounts of protein-bound phosphate: about 12 mol of phosphate per mole of titin subunit and 11 mol of phosphate per mole of nebulin subunit. Incubation of intact, excised mouse diaphragm with radioactive inorganic phosphate resulted in significant incorporation of radiophosphate into titin and nebulin. The identification of titin and nebulin phosphorylation was facilitated by a simple salt fractionation and nuclease digestion procedure that effectively separated titin and nebulin from radiolabeled nucleic acids. Such in vivo phosphorylation studies indicated that approximately 2 mol of phosphate per titin subunit and 5 to 7 mol of phosphate per nebulin subunit were incorporated within 5 h of incubation. The incorporation nearly doubled when the beta-adrenergic agonist, isoproterenol, or a phosphodiesterase inhibitor, theophylline, was present in the medium. For both proteins, phosphorylation occurred mainly on serine residues. Nebulin also appears to possess a smaller number of threonine sites. Taken together, our data indicate that a small proportion (20 to 40%) of the steady-state titin phosphates are rapidly turning over. In contrast, most of the nebulin phosphates (50 to 100%) are readily exchanged. The modulation of turnover by external stimuli that increase cytosolic cAMP raises the possibility that at least a portion of the multiple phosphorylation sites of titin and nebulin may be involved in the functional regulation of the sarcomere matrix.

Amino Acids↗
PubMed1988

Nebulin cDNAs detect a 25-kilobase transcript in skeletal muscle and localize to human chromosome 2.

By virtue of the protein's size, myofibrillar localization, and proposed functional role, the gene encoding the giant sarcomere matrix protein nebulin represents a possible site for myopathic mutations. Using polyclonal anti-nebulin antisera to screen a cDNA expression library, we have isolated and characterized two separate human fetal muscle cDNA clones. By recovering fusion polypeptide-bound portions of our polyclonal antiserum and reutilizing them to probe Western blots, we further demonstrate that the expressed cDNAs encode polypeptide epitopes unique to the protein nebulin. Both cDNAs detect a 25-kb skeletal muscle RNA transcript and localize to human chromosome 2. The identification of nebulin cDNA clones enables the complete analysis of this enormous mRNA by transcript walking through muscle cDNA libraries. Here we report a restriction map of the 3' end of the human nebulin transcript, with reference to the genomic fragments identified by the cDNA.

Chromosome Mapping↗
PubMed1988

Architecture of the sarcomere matrix of skeletal muscle: immunoelectron microscopic evidence that suggests a set of parallel inextensible nebulin filaments anchored at the Z line.

Nebulin, a giant myofibrillar protein (600-800 kD) that is abundant (3%) in the sarcomere of a wide range of skeletal muscles, has been proposed as a component of a cytoskeletal matrix that coexists with actin and myosin filaments within the sarcomere. Immunoblot analysis indicates that although polypeptides of similar size are present in cardiac and smooth muscles at low abundance, those proteins show no immunological cross-reactivity with skeletal muscle nebulin. Gel analysis reveals that nebulins in various skeletal muscles of rabbit belong to at least two classes of size variants. A monospecific antibody has been used to localize nebulin by immunoelectron microscopy in a mechanically split rabbit psoas muscle fiber preparation. Labeled split fibers exhibit six pairs of stripes of antibody-imparted transverse densities spaced at 0.1-1.0 micron from the Z line within each sarcomere. These epitopes maintain a fixed distance to the Z line irrespective of sarcomere length and do not exhibit the characteristic elastic stretch-response of titin epitopes within the I band domain. It is proposed that nebulin constitutes a set of inextensible filaments attached at one end to the Z line and that nebulin filaments are in parallel, and not in series, with titin filaments. Thus the skeletal muscle sarcomere may have two sets of nonactomyosin filaments: a set of I segment-linked nebulin filaments and a set of A segment-linked titin filaments. This four-filament sarcomere model raises the possibility that nebulin and titin might act as organizing templates and length-determining factors for actin and myosin respectively.

Actins↗
PubMed1988

Reaction 6Li

Explore the source record for details and available documents.

Journal Article↗
PubMed1988

Glycine betaine allows enhanced induction of the Agrobacterium tumefaciens vir genes by acetosyringone at low pH.

We established growth conditions for efficient induction of the vir genes of Agrobacterium tumefaciens by acetosyringone. Optimal induction was attained at a pH below 5.2 in an AB minimal medium-derived high-osmotic-strength medium containing glycine betaine. This natural osmoprotectant accelerated the adaptation of the bacteria to these conditions. We established the kinetics of induction for virB, virD, virE, and virG by using lacZ fusions, and we found that the virB mutant strain could not adapt to this low-pH medium unless 1 mM CaCl2 was added. This pH control of vir gene expression was shown to act at the level of expression of virG, which was the limiting factor. This improved vir induction at a low pH correlated with an increase in a set of proteins which was analyzed by two-dimensional gel electrophoresis. The fact that high inducibility corresponded to a reduced growth rate and the demonstration that a set of proteins was associated with the inducible state suggest that vir gene induction is linked to the adaptation of the cells to an unfavorable environment. Hence, vir gene expression in A. tumefaciens is probably dependent upon a machinery which is specific to an adaptive response; the implications for plant transformation are discussed.

Acetophenones↗
PubMed1988

Value of the yolk sac in evaluating early pregnancies.

To determine the potential value in identifying a yolk sac in women with suspected ectopic pregnancies, 211 consecutive women who were referred to pelvic sonography with this clinical indication were prospectively studied. Of the 211 patients examined, 104 proved to have a normal intrauterine pregnancy (IUP), 59 had an abnormal (nonviable) IUP, and 48 had a surgically-proven ectopic pregnancy. Considering only intrauterine "sacs" that lacked a visible embryo, a yolk sack was identified in 16 of 26 (62%) normal IUPs, and 3 of 19 (16%) abnormal IUPs, but was not seen in any of six pseudo-gestational sacs in women with ectopic pregnancies. In comparison, a double decidual sac (DDS) finding was thought to be present in 24 of 26 (92%) normal gestational sacs, 12 of 19 (63%) abnormal gestational sacs, and 2 of 6 (33%) pseudogestational sacs. We conclude that in women in whom a living embryo is not identified, demonstration of a yolk sac is reliable evidence for early IUP, and virtually excludes the possibility of an ectopic gestation. Although a yolk sac is less frequently observed than a DDS finding, it is also more specific evidence for an IUP. Careful attention to the yolk sac should permit earlier, more reliable diagnoses of IUP than previously possible.

Female↗
PubMed1987

In vivo phosphorylation of titin and nebulin in frog skeletal muscle.

Titin and nebulin are two major protein components of the sarcomere matrix in striated muscles. Purified titin and nebulin from frog (Xenopus laevis) skeletal muscle are similar in size and in amino acid composition to their mammalian or avian counterparts. Both proteins contain substantial amounts of protein-bound phosphate: about 5 to 6 per titin subunit and 8 to 9 per nebulin subunit. Injection of radioactive inorganic phosphate into the dorsal lymph sacs of Xenopus laevis resulted in the significant incorporation of radioactivity into titin and nebulin within three days of incubation. Purified titin from in vivo labeled frog gastrocnemius muscle contains one mole of radioactive phosphoserine per mole of titin subunit. These data indicate that phosphorylation of frog titin and nebulin occurs in vivo.

Amino Acids↗
PubMed1987

[Hemihepatectomy under hepato-portal interruption at normal temperature for liver malignancies--a report of 20 patients].

From July 1984 to December 1985, hemihepatectomy was done in 20 liver cancer patients under normothermic interruption of porta hepatis. There were 19 primary and 1 secondary liver carcinomas. Of the former, 17 (89%) were associated with mild or moderate cirrhosis. The peak age ranged 36-60 years. Right hemihepatectomy was performed in 18 and left hemihepatectomy in 2 with an operative mortality of 0%. Hepatic failure or secondary bleeding was not found. In the specimens resected, the largest weight was 2,500 gm. The normothermic interruption of porta hepatis usually lasted 15-25 minutes, a time long enough for hemihepatectomy. This procedure, being simple in manipulation and less detrimental to physiologic and biochemical balance in the human body, is relatively practical and beneficial to hepatectomy.

Adult↗
PubMed1985

Adenovirus sequences required for replication in vivo.

We have studied the in vivo replication properties of plasmids carrying deletion mutations within cloned adenovirus terminal sequences. Deletion mapping located the adenovirus DNA replication origin entirely within the first 67 bp of the adenovirus inverted terminal repeat. This region could be further subdivided into two functional domains: a minimal replication origin and an adjacent auxillary region which boosted the efficiency of replication by more than 100-fold. The minimal origin occupies the first 18 to 21 bp and includes sequences conserved between all adenovirus serotypes. The adjacent auxillary region extends past nucleotide 36 but not past nucleotide 67 and contains the binding site for nuclear factor I.

Adenoviridae↗
PubMed1984

Cytoskeletal matrix in striated muscle: the role of titin, nebulin and intermediate filaments.

In this chapter, first I will briefly describe the molecular properties of titin and nebulin --two extremely large, myofibrillar proteins--and discuss their distribution and organization in the sarcomere. Although these novel proteins are major myofibrillar components of a wide range of striated muscles, they have escaped the attention of muscle biochemists until very recently. As I shall point out below, biochemical studies of these proteins have been unexpectedly challenging; many standard techniques had to be modified before they became capable of handling such giant proteins. In addition, our structural studies of these proteins have encountered a challange of a different nature: how to explain their distribution in the sarcomere according to the currently accepted two filament sarcomere model, because these proteins do not appear to be thick or thin filament-associated regulatory or anchoring proteins. These studies have led us to reexamine the question of whether continuous, longitudinal filaments exist within the sarcomere of striated muscle. I will attempt to integrate our results, as well as available literature data, within the framework of a hypothetical sarcomere model which incorporates an elastic filamentous matrix consisting of titin and nebulin as additional sarcomere constituents. Finally, I will very briefly mention our recent findings that an extensive three dimensional network of intermediate (10 nm) filaments, distinct from titin and nebulin , is intimately associated with the sarcomere of adult striated muscle. I believe that the recognition of the existence of two sets of sarcomere-associated cytoskeletal filaments within adult striated muscle fibers may be a significant step toward resolving some of the unsettled questions in muscle mechanics such as those that have been discussed in this meeting.

Animals↗