Completion of the primary structure of human high-molecular-mass kininogen. The amino acid sequence of the entire heavy chain and evidence for its evolution by gene triplication

ABSTRACT The polypeptide and structural gene for a high-molecular-massc-type cytochrome, cytochromec 553O, was isolated from the methanotrophMethylococcus capsulatus Bath. Cytochromec 553O is a homodimer with a subunit molecular mass of 124,350 Da and an isoelectric point of 6.0. The hemec concentration was estimated to be 8.2 ± 0.4 mol of heme c per subunit. The electron paramagnetic resonance spectrum showed the presence of multiple low spin, S = 1/2, hemes. A degenerate oligonucleotide probe synthesized based on the N-terminal amino acid sequence of cytochrome c 553O was used to identify a DNA fragment from M. capsulatusBath that contains occ, the gene encoding cytochrome c 553O. occ is part of a gene cluster which contains three other open reading frames (ORFs). ORF1 encodes a putative periplasmic c-type cytochrome with a molecular mass of 118,620 Da that shows approximately 40% amino acid sequence identity with occand contains nine c-heme-binding motifs. ORF3 encodes a putative periplasmic c-type cytochrome with a molecular mass of 94,000 Da and contains sevenc-heme-binding motifs but shows no sequence homology toocc or ORF1. ORF4 encodes a putative 11,100-Da protein. The four ORFs have no apparent similarity to any proteins in the GenBank database. The subunit molecular masses, arrangement and number of hemes, and amino acid sequences demonstrate that cytochrome c 553O and the gene products of ORF1 and ORF3 constitute a new class of c-type cytochrome.

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A high molecular mass protein isolated from chicken gizzard: its localization at the dense plaques and dense bodies of smooth muscle and the Z-disks of skeletal muscle

Journal of Cell Science ◽

10.1242/jcs.108.3.857 ◽

1995 ◽

Vol 108 (3) ◽

pp. 857-868

Author(s):

A.G. Terasaki ◽

H. Nakagawa ◽

E. Kotani ◽

H. Mori ◽

K. Ohashi

Keyword(s):

Skeletal Muscle ◽

Smooth Muscle ◽

Amino Acid ◽

Amino Acid Sequence ◽

Molecular Mass ◽

Muscle Cells ◽

High Molecular Mass ◽

Dense Bodies ◽

Muscle Tissues ◽

Low Salt

We purified a 450 kDa protein from a low-salt alkaline extract of chicken gizzard smooth muscle. This high molecular mass protein could be extracted with the low-salt alkaline solution at 37 degrees C but not at 4 degrees C. The 450 kDa protein was isolated from the extract by ammonium sulfate fractionation and following sequential column chromatography using hydroxylapatite, DEAE-Cellulofine A-800m and phenyl-Sepharose CL-4B resins. The partially purified protein molecule resembled a flexible rod with a globular head and an irregular-shaped tail. Its length was approximately 300 nm. The nucleotide sequence of the partial cDNA encoding this protein was determined and analyzed with a data base. The analysis showed that the protein revealed significant homology with the rod region of chicken filamin (57% homology in amino acid sequence). Immunoblot analysis showed that an affinity-purified antibody reacted exclusively with the 450 kDa protein band of smooth, skeletal and cardiac muscle tissues. By indirect immunofluorescence microscopy, we examined the localization of the 450 kDa protein in smooth and skeletal muscle cells. The affinity-purified antibody against the 450 kDa protein stained the dense plaques and dense bodies of smooth muscle, the peripheral region of Z-disks and the subsarcolemmal region of skeletal muscle. Immunoelectron microscopy confirmed the localization of the 450 kDa protein at the peripheral regions of the actin anchoring structures mentioned above. Judging from its amino acid sequence, molecular size, molecular shape, immunological reactivity and localization in muscle cells, the 450 kDa protein seemed to be a new component associated with the actin-anchoring structures of muscle tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

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Characterization of a Novel Thrombin-like Enzyme, Globlase from the Venom of Gloydius blomhoffii (Japanese Mamushi)

International Journal of Biochemistry Research & Review ◽

10.9734/ijbcrr/2019/v26i430106 ◽

2019 ◽

pp. 1-8

Author(s):

Yumiko Komori ◽

Yusuke Takashima ◽

Shoko Ono ◽

Toshiaki Nikai

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Molecular Mass ◽

Isoelectric Point ◽

Primary Structure ◽

Polyacrylamide Gel Electrophoresis ◽

Hydrolytic Activity ◽

The Other ◽

Clotting Factor ◽

Human Fibrinogen

Aims: To elucidate the coagulation mechanisms of a novel clotting factor isolated from Gloydius blomhoffii venom, its hydrolytic activity on various substrates were examined. Furthermore the primary structure was determined and compared with the other snake venom components. Methodology: A thrombin-like enzyme was isolated from the crude venom of G. blomhoffii by DE52 Cellulose and CM52 Cellulose column chromatography. Enzyme activity was measured by using synthetic substrates (arginine esters, MCA-substrates and 3-(Acyloxy)-4-nitrobenzoic acid). Effect on fibrinogen was detected with bovine and human fibrinogen. Isoelectric point and molecular mass were measured by polyacrylamide gel electrophoresis and MALDI-TOF-MS. Amino acid sequence was decided with a protein sequencer by analyzing enzymatically cleaved peptides. Results: A clotting factor was found to be homologous as indicated by a single band on SDS-PAGE, and the final preparation was named as globlase. Molecular mass of this enzyme was determined to be 13,876.36 Da and the isoelectric point was 8.8. Globlase showed arginine ester hydrolytic activity, and specificity for substrates of thrombin. Proteolytic activity and phospholipase A2 (PLA2) activity were not detected. Complete amino acid sequence analysis indicated that the primary structure of globlase is similar to PLA2. However the aspartic acid which exists in the active site of PLA2 was found to be substituted by glutamine. Conclusion: It was shown in our current investigation that globlase is a novel thrombin-like enzyme isolated from G. blomhoffii venom. It was revealed that this enzyme had structure unlike the serine-protease such as the other thrombin-like enzymes.

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The amino acid sequence of the light chain of human high-molecular-mass kiniogen

European Journal of Biochemistry ◽

10.1111/j.1432-1033.1985.tb09199.x ◽

1985 ◽

Vol 152 (2) ◽

pp. 307-314 ◽

Cited By ~ 37

Author(s):

Friedrich LOTTSPEICH ◽

Josef KELLERMANN ◽

Agnes HENSCHEN ◽

Berthold FOERTSCH ◽

Werner MuLLER-ESTERL

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Molecular Mass ◽

Light Chain ◽

High Molecular Mass

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Primary structure of a human IgA1 immunoglobulin. IV. Streptococcal IgA1 protease, digestion, Fab and Fc fragments, and the complete amino acid sequence of the alpha 1 heavy chain.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)30153-9 ◽

1979 ◽

Vol 254 (8) ◽

pp. 2865-2874 ◽

Cited By ~ 3

Author(s):

F.W. Putnam ◽

Y.S. Liu ◽

T.L. Low

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Heavy Chain ◽

Primary Structure ◽

Complete Amino Acid Sequence ◽

Protease Digestion ◽

Iga1 Protease

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Phosphorylation, glycosylation and amino acid sequence of component PP3 from the proteose peptone fraction of bovine milk

Journal of Dairy Research ◽

10.1017/s0022029900027886 ◽

1993 ◽

Vol 60 (4) ◽

pp. 535-542 ◽

Cited By ~ 63

Author(s):

Esben S. Sørensen ◽

Torben E. Petersen

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Molecular Mass ◽

Primary Structure ◽

Bovine Milk ◽

Amino Sugar ◽

Amino Sugars ◽

Amino Acid Residues ◽

Proteose Peptone ◽

Amino Acid Analyser

SummaryComponent PP3 is a phosphorylated glycoprotein with an apparent molecular mass of 28 kDa isolated from the proteose peptone fraction of bovine milk. The function of the protein is not known. The primary structure has been determined and shown to contain 135 amino acid residues (EMBL accession no. P80195). It was phosphorylated at Ser29, Ser34, Ser38, Ser40 and Ser46. Two O-linked carbohydrate groups were found at Thr16 and Thr86, while one N-linked carbohydrate group was present at Asn77. Thr16 was only ∼ 50% glycosylated. The amino sugar detected by the amino acid analyser at Thr86 was mainly galactosamine but a small amount of glucosamine was also present. The amino sugars found in the carbohydrate group linked to Asn77 were both glucosamine and galactosamine. A fragment of PP3 has been isolated from milk and shown to correspond to residues 54–135. This fragment was probably generated by plasmin hydrolysing the Arg53–Ser54 bond.

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