Amino-terminal sequence analysis of the structural proteins of Sindbis virus.

The conversion of human prothrombin to thrombin is associated with a number of cleavage intermediates and products whose appearance and concentration are dependent upon the prothrombin activation conditions used. In the current investigation, the fragments of prothrombin which appear in normal human plasma after activation of the blood coagulation cascade were studied. Radioiodinated human prothrombin was added to platelet-poor relipidated normal human plasma and clotting initiated with Ca(II) and kaolin. The radiolabeled prothrombin cleavage products which formed were analyzed by polyacrylamide gel electrophoresis in the presence of dodecyl sulfate (SDS) and 2-mercaptoethanol (2-ME), A new product of prothrombin activation was observed. Its migration was more rapid than prethrombin 1 and slower than fragment 1.2. No previously identified products of prothrombin activation migrated to the same position in the gel.The previously unrecognized fragment was identified as fragment 1.2.3 as follows. Prothrombin was activated by factor Xa in the presence of Ca(II) and phospholipid. The desired product was isolated by absorption to and elution from barium citrate and by DEAE cellulose chromatography. This purified material, migrating identically with the unknown plasma product was homogeneous upon SDS gel electrophoresis with 2-ME. The amino terminal sequence of the isolated material was identical to that of prothrombin. Digestion of this material with either factor Xa or thrombin yielded as major products fragment 1.2 and fragment 1. (Fragment 2 and fragment 3 eluted from the gels under the conditions employed). Amino terminal sequence analysis of the factor Xa digestion products of the isolated material indicated three amino acid residues at each cycle. The sequences of fragment 1, fragment 2, and fragment 3 are consistent with this sequence analysis. On this basis we suggest that fragment 1.2.3 is a prominent product of prothrombin conversion to thrombin in plasma.

Amino-terminal sequence analysis of rat heart and muscle glycogen synthase: Homology to the rabbit enzyme and the implications for hormonal control

Archives of Biochemistry and Biophysics ◽

10.1016/0003-9861(89)90330-5 ◽

1989 ◽

Vol 268 (2) ◽

pp. 630-636 ◽

Cited By ~ 3

Author(s):

Stephen R. Jaspers ◽

Jill Rulfs ◽

Gary L. Johnson ◽

John E. Mole ◽

Thomas B. Miller

Keyword(s):

Sequence Analysis ◽

Muscle Glycogen ◽

Rat Heart ◽

Glycogen Synthase ◽

Hormonal Control ◽

Terminal Sequence ◽

Amino Terminal ◽

Amino Terminal Sequence

Human Luteinizing Hormone: Amino-Terminal Sequence Analysis of the β Subunit using [35S]Phenylisothiocyanate

Hoppe-Seyler´s Zeitschrift für physiologische Chemie ◽

10.1515/bchm2.1974.355.2.935 ◽

1974 ◽

Vol 355 (2) ◽

pp. 935-938 ◽

Cited By ~ 6

Author(s):

Henry T. Keutmann ◽

John W. Jacobs ◽

William H. Bishop ◽

Robert J. Ryan

Keyword(s):

Sequence Analysis ◽

Luteinizing Hormone ◽

Β Subunit ◽

Terminal Sequence ◽

Amino Terminal ◽

Amino Terminal Sequence

THROMBOSPONDIN INTERACTION WITH PLASMINOGEN

10.1055/s-0038-1643824 ◽

1987 ◽

Author(s):

Patricia DePoli ◽

Theresa Bacon-Baquley ◽

Daniel A Walz

Keyword(s):

Amino Acid ◽

Sequence Analysis ◽

Terminal Sequence ◽

Independent Manner ◽

Amino Terminal ◽

High Affinity ◽

Distinct Site ◽

Tissue Plasminogen ◽

Trimolecular Complex ◽

Amino Terminal Sequence

Platelet thrombospondin (TSP) interacts with plasminogen in a specific and saturable manner. TSP can form a trimolecular complex with histidine-rich glycoprotein and plasminogen and the plasminogen within such complexes can reportedly be activated by tissue plasminogen activator. We have studied the interaction of TSP with plasminogen using Western blotting of plasminogen, reduced plasmin and the elastase-generated fragments of plasminogen and their binding of iodinated TSP. TSP was found to specifically bind to plasminogen and the heavy (non-enzyme) chain of plasmin in a calcium-independent manner. Binding could be blocked by preincubation of the immobilized plasminogen or plasmin with an excess of unlabeled TSP. Plasminogen domains (kringles) were generated by limited eTastase proteolysis. TSP bound specifically to a single 51 kDa plasminogen fragment. The elastase-generated fragments were separated by lysine-Sepharose chromatography and their identities established by amino acid composition and amino-terminal sequence analysis. The 51 kDa plasminogen fragment bound to lysine-Sepharose and had an amino-terminal sequence corresponding to kringle 4 (K4) and a composition consistent with that of K4-K5-plasmin. TSP binding to this fragment was not blocked by the presence of an excess of the fragment K1-K2-K3, K4, nor miniplasminogen (K5-plasmin). Binding does not appear to be directly dependent upon the specific high-affinity lysine binding site of the 51 kDa fragment. Our data suggests that thrombospondin interacts with plasminogen at a single distinct site, and that this recognition site is at or near the K4-K5 contiguous region of plasminogen.

Amino-terminal sequence analysis of arachin

Journal of Protein Chemistry ◽

10.1007/bf01025174 ◽

1984 ◽

Vol 3 (4) ◽

pp. 395-401 ◽

Cited By ~ 20

Author(s):

R. Bhushan ◽

R. N. Goyal ◽

Anita Agarwal

Keyword(s):

Sequence Analysis ◽

Terminal Sequence ◽

Amino Terminal ◽

Amino Terminal Sequence

Amino-terminal sequence analysis of six cytochrome P-450 isozymes purified by monoclonal antibody directed immunopurification

Biochemical and Biophysical Research Communications ◽

10.1016/s0006-291x(84)80260-0 ◽

1984 ◽

Vol 123 (3) ◽

pp. 1201-1208 ◽

Cited By ~ 22

Author(s):

Kuo-Chi Cheng ◽

Henry C. Krutzsch ◽

Sang S. Park ◽

Preston H. Grantham ◽

Harry V. Gelboin ◽

...

Keyword(s):

Monoclonal Antibody ◽

Sequence Analysis ◽

Terminal Sequence ◽

Amino Terminal ◽

Cytochrome P 450 ◽

Amino Terminal Sequence

Simian virus 40 structural proteins: Amino-terminal sequence of the major capsid protein

Virology ◽

10.1016/0042-6822(74)90353-5 ◽

1974 ◽

Vol 60 (2) ◽

pp. 584-587 ◽

Cited By ~ 16

Author(s):

Elias Lazarides ◽

James G Files ◽

Klaus Weber

Keyword(s):

Capsid Protein ◽

Major Capsid Protein ◽

Simian Virus 40 ◽

Simian Virus ◽

Structural Proteins ◽

Terminal Sequence ◽

Amino Terminal ◽

Amino Terminal Sequence

Amino-terminal Sequence Analysis of Proteins Purified on a Nanomole Scale by Gel Electrophoresis

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)45237-x ◽

1972 ◽

Vol 247 (10) ◽

pp. 3242-3251 ◽

Cited By ~ 25

Author(s):

Alan M. Weiner ◽

Terry Platt ◽

Klaus Weber

Keyword(s):

Sequence Analysis ◽

Gel Electrophoresis ◽

Terminal Sequence ◽

Amino Terminal ◽

Amino Terminal Sequence