The expression, characterization, and crystallization of wild-type and selenomethionyl human chorionic gonadotropin (original) (raw)

Journal Article

,

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

,

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

,

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

,

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

,

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

,

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

,

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

,

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

1Department of Medicine, Columbia University, New York, New York 10032.

Search for other works by this author on:

Published:

01 February 1995

• • Article contents
  • Figures & tables
  • Video
  • Audio
  • Supplementary Data
• Cite

Cite

J W Lustbader, H Wu, S Birken, S Pollak, M A Gawinowicz Kolks, A M Pound, D Austen, W A Hendrickson, R E Canfield, The expression, characterization, and crystallization of wild-type and selenomethionyl human chorionic gonadotropin, Endocrinology, Volume 136, Issue 2, 1 February 1995, Pages 640–650, https://doi.org/10.1210/endo.136.2.7835298
Close

Navbar Search Filter Mobile Enter search term Search

Abstract

Although the glycoprotein hormone hCG was crystallized over 4 yr ago, it is only now that three-dimensional structural information is available. This manuscript reports the method for successful production of modified expressed hormone, the characteristics of the crystallized protein, and unexpected observations during the crystallization process. Two different routes of solution to the structure of hCG were followed. The first was based on the traditional method of heavy atom isomorphous replacement, and the second was the more novel method of expressing the protein with selenomethionine substituting for methionine and applying multiwavelength anomalous diffraction analysis. Selenomethionyl hCG was employed to successfully grow the crystals used for the solution of the structure of hCG after partial deglycosylation by hydrogen fluoride (HF) treatment. The selenomethionyl hCG proved to be more hydrophobic than the expressed form of native hCG. Furthermore, expressed forms of hCG that were deglycosylated by HF proved to be more intact and less susceptible to peptide bond cleavages during the crystallization process than the urinary form of HF-treated hCG studied previously. It was found that addition of reducing agent during the crystallization period was necessary for the growth of crystals of HF-treated selenomethionyl hCG suitable for diffraction studies. Growth of crystals of HF-treated expressed hCG were accelerated by the addition of dithiothreitol, but would successfully grow without reductant. HPLC analysis of the HF-treated hormones before and during the crystallization process was used to identify alterations in the molecules, including oxidation and aggregation, both of which may affect the growth of crystals.

This content is only available as a PDF.

Copyright © 1995 by The Endocrine Society

You do not currently have access to this article.

Personal account

• Sign in with email/username & password
• Get email alerts
• Save searches
• Purchase content
• Activate your purchase/trial code
• Add your ORCID iD

Get help with access

Institutional access

Access to content on Oxford Academic is often provided through institutional subscriptions and purchases. If you are a member of an institution with an active account, you may be able to access content in one of the following ways:

IP based access

Typically, access is provided across an institutional network to a range of IP addresses. This authentication occurs automatically, and it is not possible to sign out of an IP authenticated account.

Sign in through your institution

Choose this option to get remote access when outside your institution. Shibboleth/Open Athens technology is used to provide single sign-on between your institution’s website and Oxford Academic.

1. Click Sign in through your institution.
2. Select your institution from the list provided, which will take you to your institution's website to sign in.
3. When on the institution site, please use the credentials provided by your institution. Do not use an Oxford Academic personal account.
4. Following successful sign in, you will be returned to Oxford Academic.

If your institution is not listed or you cannot sign in to your institution’s website, please contact your librarian or administrator.

Sign in with a library card

Enter your library card number to sign in. If you cannot sign in, please contact your librarian.

Society Members

Society member access to a journal is achieved in one of the following ways:

Sign in through society site

Many societies offer single sign-on between the society website and Oxford Academic. If you see ‘Sign in through society site’ in the sign in pane within a journal:

1. Click Sign in through society site.
2. When on the society site, please use the credentials provided by that society. Do not use an Oxford Academic personal account.
3. Following successful sign in, you will be returned to Oxford Academic.

If you do not have a society account or have forgotten your username or password, please contact your society.

Sign in using a personal account

Some societies use Oxford Academic personal accounts to provide access to their members. See below.

Personal account

A personal account can be used to get email alerts, save searches, purchase content, and activate subscriptions.

Some societies use Oxford Academic personal accounts to provide access to their members.

Viewing your signed in accounts

Click the account icon in the top right to:

• View your signed in personal account and access account management features.
• View the institutional accounts that are providing access.

Signed in but can't access content

Oxford Academic is home to a wide variety of products. The institutional subscription may not cover the content that you are trying to access. If you believe you should have access to that content, please contact your librarian.

Institutional account management

For librarians and administrators, your personal account also provides access to institutional account management. Here you will find options to view and activate subscriptions, manage institutional settings and access options, access usage statistics, and more.

Purchase

Short-term Access

To purchase short-term access, please sign in to your personal account above.

Don't already have a personal account? Register

The expression, characterization, and crystallization of wild-type and selenomethionyl human chorionic gonadotropin - 24 Hours access

EUR €38.00

GBP £33.00

USD $41.00

Rental

This article is also available for rental through DeepDyve.

Citations

Views

Altmetric

Metrics

Total Views 23

4 Pageviews

19 PDF Downloads

Since 4/1/2017

×

Email alerts

More on this topic

Citing articles via

• Latest

• Most Read

• Most Cited

More from Oxford Academic